Modulators of Liver Receptor Homologue 1 (LRH-1) and Uses

ABSTRACT

This disclosure relates to modulators of liver receptor homologue 1 (LRH-1) and methods of managing disease and conditions related thereto. In certain embodiments, modulators are derivatives of hexahydropentalene. In certain embodiments, this disclosure relates to methods of treating or preventing cancer, diabetes, or cardiovascular disease by administering an effective amount of a hexahydropentalene derivative disclosed herein.

CROSS-REFERENCE TO RELATED APPLICATIONS

This application is a division of U.S. application Ser. No. 16/495,092filed Sep. 17, 2019, which is the National Stage of InternationalApplication No. PCT/US2018/022923 filed Mar. 16, 2018, which claims thebenefit of U.S. Provisional Application No. 62/473,036 filed Mar. 17,2017. The entirety of each of these applications is hereby incorporatedby reference for all purposes.

STATEMENT REGARDING FEDERALLY FUNDED RESEARCH

This invention was made with government support under DK0957504 andDK111171 awarded by the National Institutes of Health. The governmenthas certain rights in the invention.

BACKGROUND

Liver receptor homologue 1 (LRH-1) is a nuclear hormone receptor (NR)and acts as a transcription factor to control gene expression.Traditionally, LRH-1 was identified to be involved with cholesterolhomeostasis and early fetal development. The use of an LRH-1 agonist fortreating diabetes is reported in Lee et al. Nature, 2011, 474, 506-510.The medium-chain dietary phospholipid, dilauroyl-phosphatidylcholine(DLPC) was identified as a LRH-1 agonist. Diabetic mice fed DLPC hadimproved glucose tolerance and reduced hepatic fat accumulation, as wellas reduced quantities of circulating insulin, triglycerides and freefatty acids. The anti-diabetic effects were associated with changes inexpression of a select subset of LRH-1 target genes involved with lipidmetabolism. Importantly, the differences in health and on geneexpression by DLPC were absent in LRH-1 liver-specific conditionalknockout mice, directly implicating LRH-1 in these effects. In additionto DLPC, LRH-1 binds phosphatidyl-inositol 3,4,5-trisphosphate (PIP3),which is an important signaling lipid in diabetes.

LRH-1 is also aberrantly overexpressed in certain cancers. It isbelieved to promote tumor growth through estrogen receptor and β-cateninsignaling. See Christina et al. Liver receptor homolog-1 (LRH-1): apotential therapeutic target for cancer. Cancer Biol Ther, 2015, 16(7):997-1004. Whitby et al. report small molecule agonists of LRH-1. J MedChem 2006, 49(23):6652-5. See also Whitby et al., J Med Chem, 2011, 54,2266-2281; Busby et al. Probe Reports from the NIH Molecular LibrariesProgram, 2010, 1:1-55; Benod et al. Antagonists of nuclear receptorLRH-1. J Biol Chem, 2013, 288:19830-44, US2013/0210143, US2008/0227864,and US 2004/0038862.

Mays et al. report the crystal structures of LRH-1 bound to syntheticagonists. J Biol Chem, 2016, 291(49):25281-25291.

References cited herein are not an admission of prior art.

SUMMARY

This disclosure relates to modulators of liver receptor homologue 1(LRH-1) and methods of managing diseases and conditions related thereto.In certain embodiments, modulators are derivatives ofhexahydropentalene. In certain embodiments, this disclosure relates tomethods of treating or preventing diabetes, cancer, or cardiovasculardisease by administering an effective amount of a hexahydropentalenederivative disclosed herein to a subject in need thereof. In certainembodiments, derivatives of hexahydropentalene have the followingformula:

or salts thereof wherein the substituents are reported herein.

In certain embodiments, the disclosure contemplates pharmaceuticalcompositions comprising compounds disclosed herein or pharmaceuticallyacceptable salts thereof and pharmaceutically acceptable excipients. Incertain embodiments, the pharmaceutical products may be in the form of atablets, pills, capsules, gels, granules, or aqueous buffer solutions.

BRIEF DESCRIPTION OF THE DRAWINGS

FIG. 1A shows a scheme for the production of compounds disclosed herein.

FIG. 1B illustrates additional embodiments of this disclosure. GSK8470top left and top right, RJW100 analog lacking the hydroxyl group (named18a).

FIG. 1C shows a scheme for the production of compounds disclosed herein.

FIG. 1D shows a scheme for the production of compounds disclosed herein.

FIG. 2A illustrates additional embodiments of this disclosure.

FIG. 2B illustrates additional embodiments of this disclosure.

FIG. 3A illustrated overall structure, with α-helices shown in light andβ-sheets in slate. The Tif2 peptide is bound at the AFS. The ligand isbound at a single site in the binding pocket. Dashed line, region ofdisorder in the protein backbone that could not be modeled.

FIG. 3B shows an omit map (FO-FC, contoured at 2.5 σ) showing that asingle enantiomer of RJW100 is bound in the structure.

FIG. 3C shows superposition of RR-RJW100 with the ligand coordinatesfrom DLPC (purple, PDB 4DOS) indicating different binding mode ofRR-RJW100 compared with the PL ligands.

FIG. 3D, superposition of RR-RJW100 with the ligand coordinates fromPIP3 (PDB 4RWV) indicating different binding mode of RR-RJW100 comparedwith the PL ligands.

FIG. 3E shows DLPC expands the width at the mouth of the pocket by ˜3 Åcompared with RR-RJW100. The width was measured from Thr-341 to Asn-419(α-carbons).

FIG. 4A shows superposition of coordinates for GSK8470 (from PBD 3PLZ)and RR-RJW100.

FIG. 4B shows RR-RJW100 hydroxyl group was predicted to interact withresidues His-390 and Arg-393, but it is over 6 Å away from theseresidues in the structure.

FIG. 5A shows a close-up of the views of the binding pockets from thestructures of LRH-1 bound to RR-RJW100 depicting side chains of aminoacid residues that interact with each ligand. Residues that alsointeract with GSK8470 are shown, whereas unique interactions made byRR-RJW100 are shown in gray. Portions of the electron density maps areshown to highlight the interactions with Thr-352 through water (FO-FC,contoured to 1σ).

FIG. 5B shows a close-up views of the binding pockets from thestructures of LRH-1 bound to SR-RJW100.

FIG. 6A shows data indicating the introduction of the T352V mutation toLRH-1 ablates the stabilizing effects of RR-RJW00 and endo-RJW100.Purified LRH-1 LBD, initially bound to DLPC for homogeneity, wasincubated with either DMSO (control) or synthetic agonist dissolved inDMSO.

FIG. 6B shows data in luciferase reporter assays measuring LRH-1activity, using the SHP-luc reporter. Values have been normalized toconstitutive Renilla luciferase signal and are presented as fold changeversus wild-type LRH-1+DMSO. The A349F mutation introduces a bulkyaromatic side chain, which blocks the binding pocket and preventsbinding of synthetic ligands.

FIG. 6C shows data in luciferase reporter assays measuring LRH-1activity, using the SHP-luc reporter.

FIG. 6D shows data in luciferase reporter assays measuring LRH-1activity, using the SHP-luc reporter.

FIG. 7 shows a scheme for the production of compounds disclosed herein.

FIG. 8 shows schemes for the production of compounds disclosed herein.

DETAILED DISCUSSION

Before the present disclosure is described in greater detail, it is tobe understood that this disclosure is not limited to particularembodiments described, and as such may, of course, vary. It is also tobe understood that the terminology used herein is for the purpose ofdescribing particular embodiments only, and is not intended to belimiting, since the scope of the present disclosure will be limited onlyby the appended claims.

Unless defined otherwise, all technical and scientific terms used hereinhave the same meaning as commonly understood by one of ordinary skill inthe art to which this disclosure belongs. Although any methods andmaterials similar or equivalent to those described herein can also beused in the practice or testing of the present disclosure, the preferredmethods and materials are now described.

All publications and patents cited in this specification are hereinincorporated by reference as if each individual publication or patentwere specifically and individually indicated to be incorporated byreference and are incorporated herein by reference to disclose anddescribe the methods and/or materials in connection with which thepublications are cited. The citation of any publication is for itsdisclosure prior to the filing date and should not be construed as anadmission that the present disclosure is not entitled to antedate suchpublication by virtue of prior disclosure. Further, the dates ofpublication provided could be different from the actual publicationdates that may need to be independently confirmed.

As will be apparent to those of skill in the art upon reading thisdisclosure, each of the individual embodiments described and illustratedherein has discrete components and features which may be readilyseparated from or combined with the features of any of the other severalembodiments without departing from the scope or spirit of the presentdisclosure. Any recited method can be carried out in the order of eventsrecited or in any other order that is logically possible.

Embodiments of the present disclosure will employ, unless otherwiseindicated, techniques of medicine, organic chemistry, biochemistry,molecular biology, pharmacology, and the like, which are within theskill of the art. Such techniques are explained fully in the literature.

It must be noted that, as used in the specification and the appendedclaims, the singular forms “a,” “an,” and “the” include plural referentsunless the context clearly dictates otherwise. In this specification andin the claims that follow, reference will be made to a number of termsthat shall be defined to have the following meanings unless a contraryintention is apparent.

Certain of the compounds described herein may contain one or moreasymmetric centers and may give rise to enantiomers, diastereomers, andother stereoisomeric forms that can be defined, in terms of absolutestereochemistry at each asymmetric atom, as (R)- or (S)-. The presentchemical entities, pharmaceutical compositions and methods are meant toinclude all such possible isomers, including racemic mixtures, tautomerforms, hydrated forms, optically substantially pure forms andintermediate mixtures.

Unless otherwise stated, structures depicted herein are also meant toinclude compounds which differ only in the presence of one or moreisotopically enriched atoms. For example, compounds having the presentstructures except for the replacement or enrichment of a hydrogen bydeuterium or tritium at one or more atoms in the molecule, or thereplacement or enrichment of a carbon by ¹³C or ¹⁴C at one or more atomsin the molecule, are within the scope of this disclosure. In oneembodiment, provided herein are isotopically labeled compounds havingone or more hydrogen atoms replaced by or enriched by deuterium. In oneembodiment, provided herein are isotopically labeled compounds havingone or more hydrogen atoms replaced by or enriched by tritium. In oneembodiment, provided herein are isotopically labeled compounds havingone or more carbon atoms replaced or enriched by ¹³C. In one embodiment,provided herein are isotopically labeled compounds having one or morecarbon atoms replaced or enriched by ¹⁴C.

The disclosure also embraces isotopically labeled compounds that areidentical to those recited herein, except that one or more atoms arereplaced by an atom having an atomic mass or mass number different fromthe atomic mass or mass number usually found in nature. Examples ofisotopes that can be incorporated into disclosed compounds includeisotopes of hydrogen, carbon, nitrogen, oxygen, phosphorus, sulfur,fluorine, and chlorine, such as, e.g., ²H, ³H, ¹³C, ¹⁴C, ¹⁵N, ¹⁸O, ¹⁷O,³¹P, ³²P, ³⁵S, ¹⁸F, and ³⁶Cl, respectively. Certain isotopically labeledcompounds (e.g., those labeled with ³H and/or ¹⁴C) are useful incompound and/or substrate tissue distribution assays. Tritiated (i.e.,³H) and carbon-14 (i.e., ¹⁴C) isotopes can allow for ease of preparationand detectability. Further, substitution with heavier isotopes such asdeuterium (i.e., ²H) can afford certain therapeutic advantages resultingfrom greater metabolic stability (e.g., increased in vivo half-life orreduced dosage requirements). Isotopically labeled disclosed compoundscan generally be prepared by substituting an isotopically labeledreagent for a non-isotopically labeled reagent.

In some embodiments, provided herein are compounds that can also containunnatural proportions of atomic isotopes at one or more of atoms thatconstitute such compounds. All isotopic variations of the compounds asdisclosed herein, whether radioactive or not, are encompassed within thescope of the present disclosure.

A “linking group” refers to any variety of molecular arrangements thatcan be used to bridge to molecular moieties together. An example formulamay be —R_(m)— wherein R is selected individually and independently ateach occurrence as: —CR_(m)R_(m)—, —CHR_(m)—, —CH—, —C—, —CH₂—,—C(OH)R_(m), —C(OH)(OH)—, —C(OH)H, —C(Hal)R_(m)—, —C(Hal)(Hal)-,—C(Hal)H—, —C(N₃)R_(m)—, —C(CN)R_(m)—, —C(CN)(CN)—, —C(CN)H—,—C(N₃)(N₃)—, —C(N₃)H—, —O—, —S—, —N—, —NH—, —NR_(m)—, —(C═O)—, —(C═NH)—,—(C═S)—, —(C═CH₂)—, which may contain single, double, or triple bondsindividually and independently between the R groups. If an R is branchedwith an R_(m) it may be terminated with a group such as —CH₃, —H,—CH═CH₂, —CCH, —OH, —SH, —NH₂, —N₃, —CN, or -Hal, or two branched Rs mayform a cyclic structure. It is contemplated that in certain instances,the total Rs or “m” may be less than 100, 50, 25, 10, 5, 4, or 3.Examples of linking groups in include bridging alkyl groups, alkoxyalkylgroups, and polyethylene glycol. The term “Hal” refers to a halogen.

As used herein, a “lipid” group refers to a hydrophobic group that isnaturally or non-naturally occurring that is highly insoluble in water.As used herein a lipid group is considered highly insoluble in waterwhen the point of connection on the lipid is replaced with a hydrogenand the resulting compound has a solubility of less than 3×10⁻³ w/w (at25° C.) in water, e.g., 9.5×10⁻⁴% w/w (at 25° C.) which is the percentsolubility of hexane in water by weight. See Solvent Recovery Handbook,2d Ed, Smallwood, 2002 by Blackwell Science, page 193. Examples ofnaturally occurring lipids include saturated or unsaturated hydrocarbonchains found in fatty acids, glycerolipids, cholesterol, steroids,polyketides, and derivatives. Non-naturally occurring lipids includederivatives of naturally occurring lipids, acrylic polymers, andalkylated compounds and derivatives thereof.

As used herein, “alkyl” means a noncyclic straight chain or branched,unsaturated or saturated hydrocarbon such as those containing from 1 to22 carbon atoms, while the term “lower alkyl” or “C₁₋₄alkyl” has thesame meaning as alkyl but contains from 1 to 4 carbon atoms. The term“higher alkyl” has the same meaning as alkyl but contains from 8 to 22carbon atoms. Representative saturated straight chain alkyls includemethyl, ethyl, n-propyl, n-butyl, n-pentyl, n-hexyl, n-septyl, n-octyl,n-nonyl, and the like; while saturated branched alkyls includeisopropyl, sec-butyl, isobutyl, tert-butyl, isopentyl, and the like.Unsaturated alkyls contain at least one double or triple bond betweenadjacent carbon atoms (referred to as an “alkenyl” or “alkynyl”,respectively). Representative straight chain and branched alkenylsinclude ethylenyl, propylenyl, 1-butenyl, 2-butenyl, isobutylenyl,1-pentenyl, 2-pentenyl, 3-methyl-1-butenyl, 2-methyl-2-butenyl,2,3-dimethyl-2-butenyl, and the like; while representative straightchain and branched alkynyls include acetylenyl, propynyl, 1-butynyl,2-butynyl, 1-pentynyl, 2-pentynyl, 3-methyl-1-butynyl, and the like.

Non-aromatic mono or polycyclic alkyls are referred to herein as“carbocycles” or “carbocyclyl” groups. Representative saturatedcarbocycles include cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl,and the like; while unsaturated carbocycles include cyclopentenyl andcyclohexenyl, and the like.

“Heterocarbocycles“or heterocarbocyclyl” groups are carbocycles whichcontain from 1 to 4 heteroatoms independently selected from nitrogen,oxygen and sulfur which may be saturated or unsaturated (but notaromatic), monocyclic or polycyclic, and wherein the nitrogen and sulfurheteroatoms may be optionally oxidized, and the nitrogen heteroatom maybe optionally quaternized. Heterocarbocycles include morpholinyl,pyrrolidinonyl, pyrrolidinyl, piperidinyl, hydantoinyl, valerolactamyl,oxiranyl, oxetanyl, tetrahydrofuranyl, tetrahydropyranyl,tetrahydropyridinyl, tetrahydroprimidinyl, tetrahydrothiophenyl,tetrahydrothiopyranyl, tetrahydropyrimidinyl, tetrahydrothiophenyl,tetrahydrothiopyranyl, and the like.

“Aryl” means an aromatic carbocyclic monocyclic or polycyclic ring suchas phenyl or naphthyl. Polycyclic ring systems may, but are not requiredto, contain one or more non-aromatic rings, as long as one of the ringsis aromatic. “Arylalkyl” means an alkyl substituted with an aryl, e.g.,benzyl, methyl substituted with phenyl.

As used herein, “heteroaryl” refers to an aromatic heterocarbocyclehaving 1 to 4 heteroatoms selected from nitrogen, oxygen and sulfur, andcontaining at least 1 carbon atom, including both mono- and polycyclicring systems. Polycyclic ring systems may, but are not required to,contain one or more non-aromatic rings, as long as one of the rings isaromatic. Representative heteroaryls are furyl, benzofuranyl,thiophenyl, benzothiophenyl, pyrrolyl, indolyl, isoindolyl, azaindolyl,pyridyl, quinolinyl, isoquinolinyl, oxazolyl, isooxazolyl, benzoxazolyl,pyrazolyl, imidazolyl, benzimidazolyl, thiazolyl, benzothiazolyl,isothiazolyl, pyridazinyl, pyrimidinyl, pyrazinyl, triazinyl,cinnolinyl, phthalazinyl, and quinazolinyl. It is contemplated that theuse of the term “heteroaryl” includes N-alkylated derivatives such as a1-methylimidazol-5-yl substituent.

As used herein, “heterocycle” or “heterocyclyl” refers to mono- andpolycyclic ring systems having 1 to 4 heteroatoms selected fromnitrogen, oxygen and sulfur, and containing at least 1 carbon atom. Themono- and polycyclic ring systems may be aromatic, non-aromatic ormixtures of aromatic and non-aromatic rings. Heterocycle includesheterocarbocycles, heteroaryls, and the like.

“Alkylthio” refers to an alkyl group as defined above attached through asulfur bridge. An example of an alkylthio is methylthio, (i.e., —S—CH₃).

“Alkoxy” refers to an alkyl group as defined above attached through anoxygen bridge. Examples of alkoxy include, but are not limited to,methoxy, ethoxy, n-propoxy, i-propoxy, n-butoxy, s-butoxy, t-butoxy,n-pentoxy, and s-pentoxy. Preferred alkoxy groups are methoxy, ethoxy,n-propoxy, i-propoxy, n-butoxy, s-butoxy, and t-butoxy.

“Alkylamino” refers to an alkyl group as defined above attached throughan amino bridge. An example of an alkylamino is methylamino, (i.e.,—NH—CH₃).

“Alkanoyl” refers to an alkyl as defined above attached through acarbonyl bride (i.e., —(C═O)alkyl).

“Alkylsulfonyl” refers to an alkyl as defined above attached through asulfonyl bridge (i.e., —S(═O)₂alkyl) such as mesyl and the like,“arylsulfonyl” refers to an aryl attached through a sulfonyl bridge(i.e., —S(═O)₂aryl), and “aminosulfonyl” refers to an amino attachedthrough a sulfonyl bridge (i.e., —S(═O)₂NH₂).

“Alkylsulfinyl” refers to an alkyl as defined above attached through asulfinyl bridge (i.e. —S(═O)alkyl).

“Aminoalkyl” refers to an amino group attached through an alkyl bridge.An example of an aminoalkyl is aminomethyl, (i.e., NH₂—CH₂—).

“Hydroxyalkyl” refers to a hydroxy group attached through an alkylbridge. An example of a hydroxyalkyl is hydroxyethyl, (i.e.,HO—CH₂CH₂—).

The term “substituted” refers to a molecule wherein at least onehydrogen atom is replaced with a substituent. When substituted, one ormore of the groups are “substituents.” The molecule may be multiplysubstituted. In the case of an oxo substituent (“═O”), two hydrogenatoms are replaced. Example substituents within this context may includehalogen, hydroxy, alkyl, alkoxy, nitro, cyano, oxo, carbocyclyl,carbocycloalkyl, heterocarbocyclyl, heterocarbocycloalkyl, aryl,arylalkyl, heteroaryl, heteroarylalkyl, —NRaRb, —NRaC(═O)Rb,—NRaC(═O)NRaNRb, —NRaC(═O)ORb, —NRaSO2Rb, —C(═O)Ra, —C(═O)ORa,—C(═O)NRaRb, —OC(═O)NRaRb, —ORa, —SRa, —SORa, —S(═O)₂Ra, —OS(═O)₂Ra and—S(═O)₂Ra. Ra and Rb in this context may be the same or different andindependently hydrogen, halogen hydroxyl, alkyl, alkoxy, alkyl, amino,alkylamino, dialkylamino, carbocyclyl, carbocycloalkyl,heterocarbocyclyl, heterocarbocycloalkyl, aryl, arylalkyl, heteroaryl,heteroarylalkyl.

The term “optionally substituted,” as used herein, means thatsubstitution is optional and therefore it is possible for the designatedatom to be unsubstituted.

As used herein, “salts” refer to derivatives of the disclosed compoundswhere the parent compound is modified making acid or base salts thereof.Examples of salts include, but are not limited to, mineral salts such assodium, potassium, or zinc carboxylic acid salts, or organic acid saltsof basic residues such as amines, alkylamines, or dialkylamines; alkalior organic salts of acidic residues such as carboxylic acids; and thelike. In typical embodiments, the salts are conventional nontoxicpharmaceutically acceptable salts including the quaternary ammoniumsalts of the parent compound formed, and non-toxic inorganic or organicacids. Preferred salts include those derived from inorganic acids suchas hydrochloric, hydrobromic, sulfuric, sulfamic, phosphoric, nitric andthe like; and the salts prepared from organic acids such as acetic,propionic, succinic, glycolic, stearic, lactic, malic, tartaric, citric,ascorbic, pamoic, maleic, hydroxymaleic, phenylacetic, glutamic,benzoic, salicylic, sulfanilic, 2-acetoxybenzoic, fumaric,toluenesulfonic, methanesulfonic, ethane disulfonic, oxalic, isethionic,and the like.

“Subject” refers to any animal, preferably a human patient, livestock,rodent, monkey or domestic pet.

As used herein, the term “derivative” refers to a structurally similarcompound that retains sufficient functional attributes of the identifiedanalogue. The derivative may be structurally similar because it islacking one or more atoms, substituted with one or more substituents, asalt, in different hydration/oxidation states, e.g., substituting asingle or double bond, substituting a hydroxy group for a ketone, orbecause one or more atoms within the molecule are switched, such as, butnot limited to, replacing an oxygen atom with a sulfur or nitrogen atomor replacing an amino group with a hydroxyl group or vice versa.Replacing a carbon with nitrogen in an aromatic ring is a contemplatedderivative. The derivative may be a prodrug. Derivatives may be preparedby any variety of synthetic methods or appropriate adaptations presentedin the chemical literature or as in synthetic or organic chemistry textbooks, such as those provide in March's Advanced Organic Chemistry:Reactions, Mechanisms, and Structure, Wiley, 6th Edition (2007) MichaelB. Smith or Domino Reactions in Organic Synthesis, Wiley (2006) Lutz F.Tietze hereby incorporated by reference.

The term “prodrug” refers to an agent that is converted into abiologically active form in vivo. Prodrugs are often useful because, insome situations, they may be easier to administer than the parentcompound. They may, for instance, be bioavailable by oral administrationwhereas the parent compound is not. The prodrug may also have improvedsolubility in pharmaceutical compositions over the parent drug. Aprodrug may be converted into the parent drug by various mechanisms,including enzymatic processes and metabolic hydrolysis. Typical prodrugsare pharmaceutically acceptable esters or enol ethers. Prodrugs includecompounds wherein a hydroxy, amino or mercapto group is bonded to anygroup that, when the prodrug of the active compound is administered to asubject, cleaves to form a free hydroxy, free amino or free mercaptogroup, respectively. Examples of prodrugs include, but are not limitedto, acetate, formate and benzoate derivatives of an alcohol oracetamide, formamide and benzamide derivatives of an amine functionalgroup in the active compound and the like.

For example, if a disclosed compound or a pharmaceutically acceptableform of the compound contains a carboxylic acid functional group, aprodrug can comprise a pharmaceutically acceptable ester formed by thereplacement of the hydrogen atom of the acid group with a group such as(C₁-C₈)alkyl, (C₂-C₁₂)alkanoyloxymethyl, 1-(alkanoyloxy)ethyl havingfrom 4 to 9 carbon atoms, 1-methyl-1-(alkanoyloxy)-ethyl having from 5to 10 carbon atoms, alkoxycarbonyloxymethyl having from 3 to 6 carbonatoms, 1-(alkoxycarbonyloxy)ethyl having from 4 to 7 carbon atoms,1-methyl-1-(alkoxycarbonyloxy)ethyl having from 5 to 8 carbon atoms,N-(alkoxycarbonyl)aminomethyl having from 3 to 9 carbon atoms,1-(N-(alkoxycarbonyl)amino)ethyl having from 4 to 10 carbon atoms,3-phthalidyl, 4-crotonolactonyl, gamma-butyrolacton-4-yl,di-N,N—(C₁-C₂)alkylamino(C₂-C₃)alkyl (such as beta-dimethylaminoethyl),carbamoyl-(C₁-C₂)alkyl, N,N-di(C₁-C₂)alkylcarbamoyl-(C₁-C₂)alkyl andpiperidino-, pyrrolidino- or morpholino(C₂-C₃)alkyl.

If a disclosed compound or a pharmaceutically acceptable form of thecompound contains an alcohol functional group, a prodrug can be formedby the replacement of the hydrogen atom of the alcohol group with agroup such as (C₁-C₆)alkanoyloxymethyl, 1-((C₁-C₆)alkanoyloxy) ethyl,1-methyl-1((C₁-C₆)alkanoyloxy)ethyl (C₁-C₆)alkoxycarbonyloxymethyl,—N—(C₁-C₆)alkoxycarbonylaminomethyl, succinoyl, (C₁-C₆)alkanoyl,alpha-amino(C₁-C₄)alkanoyl, arylacyl and alpha-aminoacyl, oralpha-aminoacyl-alpha-aminoacyl, where each alpha-aminoacyl group isindependently selected from naturally occurring L-amino acids P(O)(OH)₂,—P(O)(O(C₁-C₆)alkyl)₂, and glycosyl (the radical resulting from theremoval of a hydroxyl group of the hemiacetal form of a carbohydrate).

If a disclosed compound or a pharmaceutically acceptable form of thecompound incorporates an amine functional group, a prodrug can be formedby the replacement of a hydrogen atom in the amine group with a groupsuch as R-carbonyl, RO-carbonyl, NRR′-carbonyl where R and R′ are eachindependently (C₁-C₁₀)alkyl, (C₃-C₇)cycloalkyl, benzyl, a naturalalpha-aminoacyl, —C(OH)C(O)OY₁ wherein Y¹ is H, (C₁-C₆)alkyl or benzyl,—C(OY₂)Y₃ wherein Y₂ is (C₁-C₄) alkyl and Y₃ is (C₁-C₆)alkyl,carboxy(C₁-C₆)alkyl, amino(C₁-C₄)alkyl or mono-Nordi-N,N—(C₁-C₆)alkylaminoalkyl, —C(Y₄)Y₅ wherein Y₄ is H or methyl and Y₅is mono-N— or di-N,N—(C₁-C₆)alkylamino, morpholino, piperidin-1-yl orpyrrolidin-1-yl.

As used herein, “pharmaceutically acceptable esters” include, but arenot limited to, alkyl, alkenyl, alkynyl, aryl, arylalkyl, and cycloalkylesters of acidic groups, including, but not limited to, carboxylicacids, phosphoric acids, phosphinic acids, sulfonic acids, sulfinicacids, and boronic acids.

As used herein, “pharmaceutically acceptable enol ethers” include, butare not limited to, derivatives of formula —C═C(OR) where R can beselected from alkyl, alkenyl, alkynyl, aryl, aralkyl, and cycloalkyl.Pharmaceutically acceptable enol esters include, but are not limited to,derivatives of formula —C═C(OC(O)R) where R can be selected fromhydrogen, alkyl, alkenyl, alkynyl, aryl, aralkyl, and cycloalkyl.

“Cancer” refers any of various cellular diseases with malignantneoplasms characterized by the proliferation of cells. It is notintended that the diseased cells must actually invade surrounding tissueand metastasize to new body sites. Cancer can involve any tissue of thebody and have many different forms in each body area. Within the contextof certain embodiments, whether “cancer is reduced” may be identified bya variety of diagnostic manners known to one skill in the art including,but not limited to, observation the reduction in size or number of tumormasses or if an increase of apoptosis of cancer cells observed, e.g., ifmore than a 5% increase in apoptosis of cancer cells is observed for asample compound compared to a control without the compound. It may alsobe identified by a change in relevant biomarker or gene expressionprofile, such as PSA for prostate cancer, HER2 for breast cancer, orothers.

A “chemotherapy agent,” “chemotherapeutic,” “anti-cancer agent” or thelike, refer to molecules that are recognized to aid in the treatment ofa cancer. Contemplated examples include the following molecules orderivatives such as temozolomide, carmustine, bevacizumab, procarbazine,lomustine, vincristine, gefitinib, erlotinib, cisplatin, carboplatin,oxaliplatin, 5-fluorouracil, gemcitabine, tegafur, raltitrexed,methotrexate, cytosine arabinoside, hydroxyurea, adriamycin, bleomycin,doxorubicin, daunomycin, epirubicin, idarubicin, mitomycin-C,dactinomycin, mithramycin, vinblastine, vindesine, vinorelbine,paclitaxel, taxol, docetaxel, etoposide, teniposide, amsacrine,topotecan, camptothecin, bortezomib, anagrelide, tamoxifen, toremifene,raloxifene, droloxifene, iodoxyfene, fulvestrant, bicalutamide,flutamide, nilutamide, cyproterone, goserelin, leuprorelin, buserelin,megestrol, anastrozole, letrozole, vorozole, exemestane, finasteride,marimastat, trastuzumab, cetuximab, dasatinib, imatinib, combretastatin,thalidomide, azacitidine, azathioprine, capecitabine, chlorambucil,cyclophosphamide, cytarabine, daunorubicin, doxifluridine, epothilone,irinotecan, mechlorethamine, mercaptopurine, mitoxantrone, pemetrexed,tioguanine, valrubicin and/or lenalidomide or combinations thereof suchas cyclophosphamide, methotrexate, 5-fluorouracil (CMF); doxorubicin,cyclophosphamide (AC); mustine, vincristine, procarbazine, prednisolone(MOPP); sdriamycin, bleomycin, vinblastine, dacarbazine (ABVD);cyclophosphamide, doxorubicin, vincristine, prednisolone (CHOP);bleomycin, etoposide, cisplatin (BEP); epirubicin, cisplatin,5-fluorouracil (ECF); epirubicin, cisplatin, capecitabine (ECX);methotrexate, vincristine, doxorubicin, cisplatin (MVAC).

As used herein, the terms “prevent” and “preventing” include theprevention of the recurrence, spread or onset. It is not intended thatthe present disclosure be limited to complete prevention. In someembodiments, the onset is delayed, or the severity of the disease isreduced.

As used herein, the terms “treat” and “treating” are not limited to thecase where the subject (e.g., patient) is cured and the disease iseradicated. Rather, embodiments, of the present disclosure alsocontemplate treatment that merely reduces symptoms, and/or delaysdisease progression.

As used herein, the term “combination with” when used to describeadministration with an additional treatment means that the agent may beadministered prior to, together with, or after the additional treatment,or a combination thereof.

Crystal Structures of the Nuclear Receptor, Liver Receptor Homolog 1,Bound to Synthetic Agonists Reveal a Mechanism of Activation

LRH-1 synthetic modulators are highly sought as pharmacological toolsand as potential therapeutic agents. A detailed exploration ofstructural mechanisms governing regulation of LRH-1 by synthetic ligandswas performed. See Mays et al., J Biol Chem, 2016, 291(49):25281-25291.Relative to the bacterial phospholipids (PL) LRH-1 agonist,dilauroylphosphatidyl choline (DLPC), the agonist (RR-RJW100) constrictsthe binding pocket and destabilizes portions of the activation functionsurface (AFS) (FIG. 1). Stabilization of the AFS may facilitateco-activator binding, leading to greater potency or efficacy.Alternatively, analogs designed to enhance the AFS destabilization maybe effective antagonists or inverse agonists. RJW100 was an effectivederivative but modestly increased LRH-1 activation.

Crystal structure experiments reveals a different binding mode forRR-RJW100 compared with GSK8470 (FIG. 1). Although this was surprising,it is not unreasonable, considering that LRH-1 has a very largehydrophobic binding pocket and that these agonists are also quitehydrophobic, filling only 37% of the available space (excluding waters).It is possible that many of the GSK8470 analogs investigated in theprevious structure-activity relationship study adopt a variety ofdifferent conformations. This seems to be the case in our dockingstudies with these ligands; multiple very different binding modes withsimilar energies are predicted. Importantly, however, the repositioningof RR-RJW100 in our structure appears to be driven by particularinteractions, because SR-RJW100 assumes a very similar pose (FIG. 1).This occurs despite the fact that the SR derivative exhibits signs ofmotion in our crystal structure, with significant disorder in the tailof the ligand and higher relative B-factors than RR-RJW100.

A major factor driving repositioning of the RJW100 isomers was thehydrogen bonding interaction made by the hydroxyl group. Although thecontact with residue Thr-352 is indirect, it is mediated by a watermolecule. The existence of conserved water molecules, as well as theirparticipation in ligand binding, has been described. Thus, thisinteraction could serve as an anchor point to secure the compound in apredictable orientation, enabling the targeting of desired parts of thebinding pocket via strategic addition of substituents to the ligand'sscaffold. Moreover, replacing the RJW100 hydroxyl group with a largerpolar moiety may allow direct contact with Thr-352, leading to astronger interaction.

The role of the Thr-352 interaction in LRH-1 activation by RR-RJW100 wasdemonstrated through the marked loss of activation by this compound whenthis residue was mutated. In addition, an RJW100 analog lacking ahydroxyl group was a poor activator. Unexpectedly, the T352V mutationalso resulted in a loss of activity for GSK8470, although this compounddoes not interact with the Thr-352-coordinated water molecule. However,we show that the T352V mutation weakens GSK8470's interaction withHis-390, perhaps via destabilization of the conserved water network.This could be responsible for the loss of activity of GSK8470 whenThr-352 is mutated.

Studies indicate that the interaction of small molecule agonists withLRH-1 is complex. Not only do these agonists affect receptorconformation differently from PL ligands, but they also exhibit anunexpected variability in binding modes.

Hexahydropentalene Derivatives

In certain embodiments, this disclosure relates to compounds that arehexahydropentalene derivatives. In certain embodiments, thehexahydropentalene derivatives are any of the compounds disclosed hereinoptionally substituted with one or more substituents. In certainembodiments, the compounds have the following formula:

including prodrugs, or salts thereof wherein,

R¹ is alkyl, halogen, nitro, cyano, hydroxy, amino, mercapto, formyl,carboxy, carbamoyl, alkoxy, hydroxyalkyl, alkylthio, thioalkyl,alkylamino, aminoalkyl, (alkyl)₂amino, alkanoyl, alkoxycarbonyl,alkylsulfinyl, alkylsulfonyl, arylsulfonyl, carbocyclyl, benozyl,benzyl, aryl, or heterocyclyl, wherein R is optionally substituted withone or more, the same or different, R¹⁰;

R² is hydrogen, alkyl, halogen, nitro, cyano, hydroxy, amino, mercapto,formyl, carboxy, carbamoyl, alkoxy, hydroxyalkyl, alkylthio, thioalkyl,alkylamino, aminoalkyl, (alkyl)₂amino, alkanoyl, alkoxycarbonyl,alkylsulfinyl, alkylsulfonyl, arylsulfonyl, carbocyclyl, benozyl,benzyl, aryl, or heterocyclyl, wherein R² is optionally substituted withone or more, the same or different, R¹⁰;

R³ is hydrogen, alkyl, halogen, nitro, cyano, hydroxy, amino, mercapto,formyl, carboxy, carbamoyl, alkoxy, hydroxyalkyl, alkylthio, thioalkyl,alkylamino, aminoalkyl, (alkyl)₂amino, alkanoyl, alkoxycarbonyl,alkylsulfinyl, alkylsulfonyl, arylsulfonyl, carbocyclyl, benozyl,benzyl, aryl, or heterocyclyl, wherein R³ is optionally substituted withone or more, the same or different, R¹⁰;

R⁴ is alkyl, halogen, nitro, cyano, hydroxy, amino, mercapto, formyl,carboxy, carbamoyl, alkoxy, hydroxyalkyl, alkylthio, thioalkyl,alkylamino, aminoalkyl, (alkyl)₂amino, alkanoyl, alkoxycarbonyl,alkylsulfinyl, alkylsulfonyl, arylsulfonyl, carbocyclyl, benozyl,benzyl, aryl, or heterocyclyl, wherein R⁴ is optionally substituted withone or more, the same or different, R¹⁰;

R⁵ is alkyl, halogen, nitro, cyano, hydroxy, amino, mercapto, formyl,carboxy, carbamoyl, alkoxy, hydroxyalkyl, alkylthio, thioalkyl,alkylamino, aminoalkyl, (alkyl)₂amino, alkanoyl, alkoxycarbonyl,alkylsulfinyl, alkylsulfonyl, arylsulfonyl, carbocyclyl, benozyl,benzyl, aryl, or heterocyclyl, wherein R⁵ is optionally substituted withone or more, the same or different, R¹⁰;

R⁶ is a lipid or alkyl wherein R⁶ is optionally terminally substitutedwith a hydroxy, carboxy, or phosphate, wherein the hydroxy, carboxy, orphosphate are optionally further substituted with R¹⁰;

R⁷ is hydrogen, alkyl, halogen, nitro, cyano, hydroxy, amino, mercapto,formyl, carboxy, carbamoyl, alkoxy, hydroxyalkyl, alkylthio, thioalkyl,alkylamino, aminoalkyl, (alkyl)₂amino, alkanoyl, alkoxycarbonyl,alkylsulfinyl, alkylsulfonyl, arylsulfonyl, carbocyclyl, benozyl,benzyl, aryl, or heterocyclyl, wherein R⁷ is optionally substituted withone or more, the same or different, R¹⁰; or

R¹ and R⁷ together are an oxo or oxime, wherein the oxime is optionallysubstituted with one or more, the same or different, R¹⁰;

R¹⁰ is alkyl, halogen, nitro, cyano, hydroxy, amino, mercapto, formyl,carboxy, carbamoyl, alkoxy, hydroxyalkyl, alkylthio, thioalkyl,alkylamino, aminoalkyl, (alkyl)₂amino, alkanoyl, alkoxycarbonyl,alkylsulfinyl, alkylsulfonyl, arylsulfonyl, aminosulfonyl, carbocyclyl,benozyl, benzyl, aryl, or heterocyclyl, wherein R¹⁰ is optionallysubstituted with one or more, the same or different, R¹¹; and

R¹¹ is halogen, nitro, cyano, hydroxy, trifluoromethoxy,trifluoromethyl, amino, formyl, carboxy, carbamoyl, mercapto, sulfamoyl,methyl, ethyl, methoxy, ethoxy, isopropoxy, tert-butoxy, hydoxymethyl,hydroxyethyl, thiomethyl, thioethyl, aminomethyl, aminoethyl, acetyl,acetoxy, methylamino, ethylamino, dimethylamino, diethylamino,N-methyl-N-ethylamino, acetylamino, N-methylcarbamoyl, N-ethylcarbamoyl,N,N-dimethylcarbamoyl, N,N-diethylcarbamoyl, N-methyl-N-ethylcarbamoyl,methylthio, ethylthio, methylsulfinyl, ethylsulfinyl, mesyl,ethylsulfonyl, methoxycarbonyl, ethoxycarbonyl, isopropoxycarbonyl,tert-butoxycarbonyl, N-methylsulfamoyl, N-ethylsulfamoyl,N,N-dimethylsulfamoyl, N,N-diethylsulfamoyl, N-methyl-N-ethylsulfamoyl,benozyl, benzyl, carbocyclyl, aryl, or heterocyclyl.

In certain embodiments, R² and R³ are hydrogen, R⁴ is 1-phenylvinyl or1-phenylethyl, and R⁵ is phenyl.

In certain embodiments, R⁶ is alkyl terminally substituted with ahydroxy, carboxy, or phosphate, wherein the hydroxy, carboxy, orphosphate are optionally further substituted with R¹⁰.

In certain embodiments, compounds have one of the following formula

wherein R⁶ is hydrogen, alkyl, or alkanoyl optionally substituted withR¹⁰.

In certain embodiments, R¹ is hydroxyl, alkyl, amino, aminoalkyl,carbamoyl, sulphate, sulfonate, aminosulfonyl, phosphate, phosphonate,or heterocyclyl, wherein R¹ is optionally substituted with R¹⁰.

In certain embodiments, R¹ is thiazolidinedione or triazole.

In certain embodiments, compounds have one of the following formula:

wherein,

X is a linking group, —CH₂—, —C(OH)(OH)—, —C(OH)H, —C(Hal)(Hal)-,—C(Hal)H—, —O—, —S—, —(S═O)—, —SO₂—, —NH—, —(C═O)—, —(C═NH)—, or—(C═S)—;

Y is —CH₂—, —C(OH)(OH)—, —C(OH)H, —C(Hal)(Hal)-, —C(Hal)H—, —O—, —S—,—(S═O)—, —SO₂—, —NH—, —(C═O)—, —(C═NH)—, or —(C═S)—;

Z is —CH₂—, —C(OH)(OH)—, —C(OH)H, —C(Hal)(Hal)-, —C(Hal)H—, —O—, —S—,—(S═O)—, —SO₂—, —NH—, —(C═O)—, —(C═NH)—, or —(C═S)—; and

R¹ is hydrogen, hydroxy, alkyl, alkanoyl, amino, aminoalkyl, carbamoyl,sulfate, sulfonate, aminosulfonyl, phosphate, phosphonate, orheterocyclyl.

In certain embodiments, X, Y, Z, and R¹ may be:

a) X is O, and R¹ is alkanoyl;

b) X is —NH—, and R¹ is alkanoyl;

c) X is O, and R¹ is aminosulfonyl;

d) X is —NH—, and R¹ is aminosulfonyl;

e) X is —(C═O)—, R¹ is amino;

f) X is O, Y is —(C═O)—, R¹ is amino;

g) X is O, Y is —(C═O)—, Z is —NH—, and R¹ is sulfonate; and

h) X is O, Y is —(C═O)—, Z is —NH—, and R¹ is aminosulfonyl.

In certain embodiments, R⁷ is hydroxyl or alkoxy, wherein R⁷ isoptionally substituted with one or more, the same or different, R¹⁰.

In certain embodiments, the compounds have the following formula:

including prodrugs, or salts thereof wherein,

R¹ is hydrogen, alkyl, halogen, nitro, cyano, hydroxy, amino, mercapto,formyl, carboxy, carbamoyl, alkoxy, hydroxyalkyl, alkylthio, thioalkyl,alkylamino, aminoalkyl, (alkyl)₂amino, alkanoyl, alkoxycarbonyl,alkylsulfinyl, alkylsulfonyl, arylsulfonyl, carbocyclyl, benozyl,benzyl, aryl, or heterocyclyl, wherein R¹ is optionally substituted withone or more, the same or different, R¹⁰;

R² is hydrogen, alkyl, halogen, nitro, cyano, hydroxy, amino, mercapto,formyl, carboxy, carbamoyl, alkoxy, hydroxyalkyl, alkylthio, thioalkyl,alkylamino, aminoalkyl, (alkyl)₂amino, alkanoyl, alkoxycarbonyl,alkylsulfinyl, alkylsulfonyl, arylsulfonyl, carbocyclyl, benozyl,benzyl, aryl, or heterocyclyl, wherein R² is optionally substituted withone or more, the same or different, R¹⁰;

R³ is hydrogen, alkyl, halogen, nitro, cyano, hydroxy, amino, mercapto,formyl, carboxy, carbamoyl, alkoxy, hydroxyalkyl, alkylthio, thioalkyl,alkylamino, aminoalkyl, (alkyl)₂amino, alkanoyl, alkoxycarbonyl,alkylsulfinyl, alkylsulfonyl, arylsulfonyl, carbocyclyl, benozyl,benzyl, aryl, or heterocyclyl, wherein R³ is optionally substituted withone or more, the same or different, R¹⁰;

R⁴ is alkyl, halogen, nitro, cyano, hydroxy, amino, mercapto, formyl,carboxy, carbamoyl, alkoxy, hydroxyalkyl, alkylthio, thioalkyl,alkylamino, aminoalkyl, (alkyl)₂amino, alkanoyl, alkoxycarbonyl,alkylsulfinyl, alkylsulfonyl, arylsulfonyl, carbocyclyl, benozyl,benzyl, aryl, or heterocyclyl, wherein R⁴ is optionally substituted withone or more, the same or different, R¹⁰;

R⁵ is alkyl, halogen, nitro, cyano, hydroxy, amino, mercapto, formyl,carboxy, carbamoyl, alkoxy, hydroxyalkyl, alkylthio, thioalkyl,alkylamino, aminoalkyl, (alkyl)₂amino, alkanoyl, alkoxycarbonyl,alkylsulfinyl, alkylsulfonyl, arylsulfonyl, carbocyclyl, benozyl,benzyl, aryl, or heterocyclyl, wherein R⁵ is optionally substituted withone or more, the same or different, R¹⁰;

R⁶ is a lipid or alkyl wherein R⁶ is optionally terminally substitutedwith a hydroxy, carboxy, or phosphate, wherein the hydroxy, carboxy, orphosphate are optionally further substituted with R¹⁰;

R⁷ is hydrogen, alkyl, halogen, nitro, cyano, hydroxy, amino, mercapto,formyl, carboxy, carbamoyl, alkoxy, hydroxyalkyl, alkylthio, thioalkyl,alkylamino, aminoalkyl, (alkyl)₂amino, alkanoyl, alkoxycarbonyl,alkylsulfinyl, alkylsulfonyl, arylsulfonyl, carbocyclyl, benozyl,benzyl, aryl, or heterocyclyl, wherein R⁷ is optionally substituted withone or more, the same or different, R¹⁰; or

R¹ and R⁷ together are an oxo or oxime, wherein the oxime is optionallysubstituted with one or more, the same or different, R¹⁰;

R¹⁰ is alkyl, halogen, nitro, cyano, hydroxy, amino, mercapto, formyl,carboxy, carbamoyl, alkoxy, hydroxyalkyl, alkylthio, thioalkyl,alkylamino, aminoalkyl, (alkyl)₂amino, alkanoyl, alkoxycarbonyl,alkylsulfinyl, alkylsulfonyl, arylsulfonyl, aminosulfonyl, carbocyclyl,benozyl, benzyl, aryl, or heterocyclyl, wherein R¹⁰ is optionallysubstituted with one or more, the same or different, R¹¹; and

R¹¹ is halogen, nitro, cyano, hydroxy, trifluoromethoxy,trifluoromethyl, amino, formyl, carboxy, carbamoyl, mercapto, sulfamoyl,methyl, ethyl, methoxy, ethoxy, isopropoxy, tert-butoxy, hydoxymethyl,hydroxyethyl, thiomethyl, thioethyl, aminomethyl, aminoethyl, acetyl,acetoxy, methylamino, ethylamino, dimethylamino, diethylamino,N-methyl-N-ethylamino, acetylamino, N-methylcarbamoyl, N-ethylcarbamoyl,N,N-dimethylcarbamoyl, N,N-diethylcarbamoyl, N-methyl-N-ethylcarbamoyl,methylthio, ethylthio, methylsulfinyl, ethylsulfinyl, mesyl,ethylsulfonyl, methoxycarbonyl, ethoxycarbonyl, isopropoxycarbonyl,tert-butoxycarbonyl, N-methylsulfamoyl, N-ethylsulfamoyl,N,N-dimethylsulfamoyl, N,N-diethylsulfamoyl, N-methyl-N-ethylsulfamoyl,benozyl, benzyl, carbocyclyl, aryl, or heterocyclyl.

R⁴ is 1-phenylvinyl or 1-phenylethyl, and R⁵ is phenyl optionallysubstituted.

In certain embodiments, R⁶ is alkyl terminally substituted with ahydroxy, carboxy, or phosphate, wherein the hydroxy, carboxy, orphosphate are optionally further substituted with R¹⁰.

Methods of Use

In certain embodiments, this disclosure relates to methods of treatingor preventing diseases or conditions associated with LRH-1 such asdiabetes, cancer, or cardiovascular disease by administering aneffective amount of a hexahydropentalene derivative disclosed herein toa subject in need thereof.

In certain embodiments, the disclosure relates to methods of treating orpreventing diabetes comprising administering an effective amount of apharmaceutical composition comprising compounds disclosed herein to asubject in need thereof. In certain embodiments, the subject is at riskof, exhibiting symptoms of, or diagnosed with diabetes,insulin-dependent diabetes mellitus, non insulin-dependent diabetesmellitus, or gestational diabetes.

In certain embodiments, this disclosure relates to compounds disclosedherein that are LRH-1 agonists for use in the prevention of progressiveloss of pancreatic beta-cells. It also relates to an LRH-1 agonist foruse in the preservation or restoration of pancreatic beta-cells.Further, it relates to an LRH-1 agonist for use in the prevention ortreatment of type I diabetes or insulin-dependent diabetes mellitus, theincrement of survival of pancreatic beta-cells, the increment of theperformance of pancreatic beta-cells, the increment of the survival of abeta-cell graft, the in vitro preservation of pancreatic beta-cells,maintaining insulin secretion and/or in a method of transplantingpancreatic islet cells.

Diabetes mellitus (DM) is often simply referred to as diabetes. Diabetesis a condition in which a person has a high blood sugar (glucose) levelas a result of the body either not producing enough insulin, or becausebody cells do not properly respond to the insulin that is produced.

In healthy persons, blood glucose levels are maintained within a narrowrange, primarily by the actions of the hormone insulin. Insulin isreleased by pancreatic beta-cells at an appropriate rate in response tocirculating glucose concentrations, the response being modulated byother factors including other circulating nutrients, islet innervationand incretin hormones. Insulin maintains glucose concentrations byconstraining the rate of hepatic glucose release to match the rate ofglucose clearance.

Insulin thus enables body cells to absorb glucose, to turn into energy.If the body cells do not absorb the glucose, the glucose accumulates inthe blood (hyperglycemia), leading to various potential medicalcomplications. Accordingly, diabetes is characterized by increased bloodglucose resulting in secondary complications such as cardiovasculardiseases, kidney failure, retinopathy and neuropathy if not properlycontrolled. Two major pathophysiologies are related to increaseglycemia. The first is an autoimmune attack against the pancreaticinsulin-producing beta-cells (Type 1 diabetes or insulin-dependentdiabetes) whilst the second is associated to poor beta-cell function andincreased peripheral insulin resistance (Type 2 diabetes or non-insulindependent diabetes). Similar to Type 1, beta-cell death is also observedin Type 2 diabetes. Type 1 and often Type 2 diabetes requires the personto inject insulin.

Type 1 DM is typically characterized by loss of the insulin-producingbeta-cells of the islets of Langerhans in the pancreas leading toinsulin deficiency. This type of diabetes can be further classified asimmune-mediated or idiopathic. The majority of Type 1 diabetes is of theimmune-mediated nature, where beta-cell loss is a T-cell mediatedautoimmune attack. Sensitivity and responsiveness to insulin are usuallynormal, especially in the early stages. Type 1 diabetes can affectchildren or adults but was traditionally termed “juvenile diabetes”because it represents a majority of the diabetes cases in children.

Type 2 DM is characterized by beta-cell dysfunction in combination withinsulin resistance. The defective responsiveness of body tissues toinsulin is believed to involve the insulin receptor. Similar to Type 1diabetes an insufficient beta cell mass is also a pathogenic factor inmany Type 2 diabetic patients. In the early stage of Type 2 diabetes,hyperglycemia can be reversed by a variety of measures and medicationsthat improve insulin secretion and reduce glucose production by theliver. As the disease progresses, the impairment of insulin secretionoccurs, and therapeutic replacement of insulin may sometimes becomenecessary in certain patients. In certain embodiments, the treatment ofdiabetes by administering compounds disclosed herein is in combinationwith the administration of insulin.

Diabetes without proper treatments can cause many complications. Acutecomplications include hyperglycaemia, diabetic ketoacidosis, ornonketotic hyperosmolar coma. Serious long-term complications includecardiovascular disease, chronic renal failure, retinal damage.

In certain embodiments, this disclosure relates to methods of treatingor preventing cardiovascular disease comprising administering aneffective amount of a pharmaceutical composition comprising a compounddisclosed herein to a subject in need thereof.

In certain embodiments, the cardiovascular disease is coronary arterydiseases (CAD), angina, myocardial infarction, stroke, hypertensiveheart disease, rheumatic heart disease, cardiomyopathy, heartarrhythmia, congenital heart disease, valvular heart disease, carditis,aortic aneurysms, peripheral artery disease, and venous thrombosis.

In certain embodiments, this disclosure relates to methods of managingcancer. In certain embodiments, this disclosure relates to methods oftreating cancer comprising administering an effective amount of an agentto a subject in need thereof. In certain embodiments, the cancer ispancreatic cancer, breast cancer, liver cancer, colon cancer, orgastrointestinal tumors.

Benod et al. report LRH-1 regulates pancreatic cancer cell growth andproliferation. Proc Natl Acad Sci USA, 2011, 108(41):16927-31. Pan etal. report LRH-1-dependent programming of mitochondrial glutamineprocessing drives liver cancer. Genes Dev, 2016, 30(11): 1255-1260.Holly et al. LRH-1 drives colon cancer cell growth by repressing theexpression of the CDKN1A gene in a p53-dependent manner. Nucleic AcidsRes, 2016, 44(2): 582-594.

In certain embodiments, the cancer is selected from bladder cancer,breast cancer, colon cancer, rectal cancer, endometrial cancer, kidneycancer, leukemia, lung cancer, melanoma, non-Hodgkin lymphoma,pancreatic cancer, prostate cancer, and thyroid cancer.

The compounds disclosed herein can be used alone in the treatment ofeach of the foregoing conditions or can be used to provide additive orpotentially synergistic effects with certain existing chemotherapies,radiation, biological or immunotherapeutics (including monoclonalantibodies) and vaccines. The compounds disclosed herein may be usefulfor restoring effectiveness of certain existing chemotherapies andradiation and or increasing sensitivity to certain existingchemotherapies and/or radiation.

Coste et al. report LRH-1-mediated glucocorticoid synthesis inenterocytes protects against inflammatory bowel disease. PNAS, 2007. 104(32) 13098-13103. See also Fernandez-Marcos et al. Emerging actions ofthe nuclear receptor LRH-1 in the gut, Biochim Biophys Acta. 2011August; 1812(8): 947-955. Mueller et al. The nuclear receptor LRH-1critically regulates extra-adrenal glucocorticoid synthesis in theintestine, Journal of Experimental Medicine September 2006, 203 (9)2057-2062.

Thus, in certain embodiments, this disclosure relates to methods toprevent or treat gut, intestinal, and colonic inflammation, comprisingadministrating a compound disclosed herein in an effective amount to asubject in need thereof. In certain embodiments, the subject is at riskof, exhibiting symptoms of, or diagnosed with intestinal and colonicinflammation.

In another aspect, the disclosure relates to methods to prevent or treatinflammatory bowel diseases (IBD), comprising administrating a compounddisclosed herein in an effective amount to a subject in need thereof. Incertain embodiments, the subject is at risk of, exhibiting symptoms of,or diagnosed with inflammatory bowel diseases (IBD).

In another aspect, the disclosure relates to methods to prevent or treatCrohn's disease, comprising administrating a compound disclosed hereinin an effective amount to a subject in need thereof. In certainembodiments, the subject is at risk of, exhibiting symptoms of, ordiagnosed with Crohn's disease.

In another aspect, the disclosure relates to methods to prevent or treatcolitis or ulcerative colitis, comprising administrating a compounddisclosed herein in an effective amount to a subject in need thereof. Incertain embodiments, the subject is at risk of, exhibiting symptoms of,or diagnosed with colitis or ulcerative colitis.

Overweight and obesity are increasingly common conditions in the world.Doctors measure body mass index (BMI) to screen for obesity. Obesity isa serious medical condition that can cause complications such asmetabolic syndrome, high blood pressure, atherosclerosis, heart disease,diabetes, high serum cholesterol, cancers and sleep disorders. Thus,there is a need to reduce obesity.

Fatty liver, or hepatic steatosis, is a term that describes the buildupof fat in the liver. Excessive alcohol use causes fat to accumulate,damages the liver, and cirrhosis may develop. Nonalcoholic fatty liverdisease (NAFLD) is a fatty liver disease associated with obesity-relateddisorders, such as type-2 diabetes and metabolic syndrome, occurring inpeople who drink little or no alcohol. Nonalcoholic steatohepatitis(NASH) is a more advanced and severe subtype of NAFLD where steatosis iscomplicated by liver-cell injury and inflammation, with or withoutfibrosis. NASH can be severe and can lead to cirrhosis, in which theliver is permanently damaged and scarred and no longer able to workproperly. Insulin resistance, altered lipid storage and metabolism,accumulation of cholesterol within the liver, oxidative stress resultingin increased hepatic injury, and bacterial translocation secondary todisruption of gut microbiota have all been implicated as importantco-factors contributing to progression of NASH. Due to the growingepidemic of obesity and diabetes, NASH is projected to become the mostcommon cause of advanced liver disease and the most common indicationfor liver transplantation.

Lee et al. report dilauroyl phosphatidylcholine (DLPC) is an LRH-1agonist ligand in vitro. DLPC treatment induces bile acid biosyntheticenzymes in mouse liver, increases bile acid levels, and lowers hepatictriglycerides and serum glucose. DLPC treatment also decreases hepaticsteatosis and improves glucose homeostasis in two mouse models ofinsulin resistance. Nature volume 474, pages 506-510 (2011). Sahini etal. report differentially expressed genes (DEGs) were identified whichare mechanistically linked to lipid droplet (LD) formation inhepatocytes. LD-associated DEGs frequently regulated in patient sampleswere identified. Liver-receptor homolog-1 (NR5A2), was commonlyrepressed among patients examined. Translational Research, 177: 41-69(2016).

In certain embodiments, this disclosure relates to methods to prevent ortreat hepatic steatosis or metabolic syndrome, comprising administratingcompounds disclosed herein in an effective amount to a subject in needthereof. In certain embodiments, the subject is at risk of, exhibitingsymptoms of, or diagnosed with nonalcoholic fatty liver disease (NAFLD).In certain embodiments, the subject is at risk of, exhibiting symptomsof, or diagnosed with nonalcoholic steatohepatitis (NASH). In certainembodiments, the subject is at risk of, exhibiting symptoms of, ordiagnosed with alcoholic liver disease (ALD). In certain embodiments,the subject is at risk of, exhibiting symptoms of, or diagnosed withalcoholic steatohepatitis (ASH).

In certain embodiments, a subject is at risk of NAFLD due to obesity,insulin resistance, an enlarged liver, signs of cirrhosis, or abnormallevels of liver enzymes, triglycerides and/or cholesterol. Signs ofinsulin resistance include darkened skin patches over your knuckles,elbows, and knees. Signs of cirrhosis include jaundice, a condition thatcauses your skin and whites of your eyes to turn yellow. A sign of NAFLDor NASH includes blood test showing increased levels of the liverenzymes alanine aminotransferase (ALT) and aspartate aminotransferase(AST). An enlarged liver or an abnormal amount of fat in a liver may beidentified by ultrasound, computerized tomography (CT) scans, magneticresonance imaging or combinations thereof. A liver biopsy may be used todetect liver inflammation and damage to diagnose NASH.

Metabolic syndrome is typically diagnosed in the presence of three ormore of the following medical issues: large waste size, e.g., 40 inchesor more, high triglycerides e.g., triglyceride level of 150 mg/dL orhigher, low levels of HDL cholesterol less than 50 mg/dL, high bloodpressure, e.g., 130/85 mmHg or higher, and high blood glucose (or bloodsugar) levels, a fasting blood sugar level of 100 mg/dL or higher.

In another aspect, the disclosure relates to methods to control orreduce the serum cholesterol level, comprising administrating compoundsdisclosed herein in an effective amount to a subject in need thereof. Incertain embodiments, the subject has a borderline high serum cholesterollevel, 200-239 mg/dL. In certain embodiments, the subject has a highserum cholesterol level, >240 mg/dL. In certain embodiments, the subjectis at risk of, exhibiting symptoms, or diagnosed withhypercholesterolemia.

In another aspect, the disclosure relates to methods to prevent or treathepatic steatosis, comprising administrating a compound disclosed hereinin an effective amount to a subject in need thereof. In certainembodiments, the subject is at risk of, exhibiting symptoms of, ordiagnosed with alcoholic liver disease (ALD). In certain embodiments,the subject is at risk of, exhibiting symptoms of, or diagnosed withalcoholic steatohepatitis (ASH). In certain embodiments, the subject isat risk of, exhibiting symptoms of, or diagnosed with nonalcoholic fattyliver disease (NAFLD). In certain embodiments, the subject is at riskof, exhibiting symptoms of, or diagnosed with nonalcoholicsteatohepatitis (NASH). In certain embodiments, a subject is at risk ofNAFLD due to obesity, insulin resistance, an enlarged liver, signs ofcirrhosis, or abnormal levels of liver enzymes, triglycerides and/orcholesterol. Signs of insulin resistance include darkened skin patchesover your knuckles, elbows, and knees. Signs of cirrhosis includejaundice, a condition that causes your skin and whites of your eyes toturn yellow. A sign of NAFLD or NASH includes blood test showingincreased levels of the liver enzymes alanine aminotransferase (ALT) andaspartate aminotransferase (AST). An enlarged liver or an abnormalamount of fat in a liver may be identified by ultrasound, computerizedtomography (CT) scans, magnetic resonance imaging or combinationsthereof. A liver biopsy may be used to detect liver inflammation anddamage to diagnose NASH.

The precise therapeutically effective amount of the compounds of thisdisclosure will depend on a number of factors. There are variablesinherent to the compounds including, but not limited to, the following:molecular weight, absorption, bioavailability, distribution in the body,tissue penetration, half-life, metabolism, protein binding, andexcretion. These variables determine what dose of compound needs to beadministered in a sufficient percentage and for a sufficient amount oftime to have the desired effect on the condition being treated (e.g.,neoplasm). The duration of drug exposure will be limited only by thecompound half-life, and side effects from treatment requiring cessationof dosing. The amount of compound administered will also depend onfactors related to patients and disease including, but not limited to,the following: the age, weight, concomitant medications and medicalcondition of the subject being treated, the precise condition requiringtreatment and its severity, the nature of the formulation, and the routeof administration. Ultimately the dose will be at the discretion of theattendant physician or veterinarian. Typically, the compound disclosedherein will be given for treatment in the range of 0.01 to 30 mg/kg bodyweight of recipient (mammal) per day or per dose or per cycle oftreatment and more usually in the range of 0.1 to 10 mg/kg body weightper day or per dose or per cycle of treatment. Thus, for an adult humanbeing treated for a condition, the actual amount per day or per dose orper cycle of treatment would usually be from 1 to 2000 mg and thisamount may be given in a single or multiple doses per day or per dose orper cycle of treatment. Dosing regimens may vary significantly and willbe determined and altered based on clinical experience with thecompound. The full spectrum of dosing regimens may be employed rangingfrom continuous dosing (with daily doses) to intermittent dosing. Atherapeutically effective amount of a pharmaceutically acceptable saltof a compound disclosed herein may be determined as a proportion of thetherapeutically effective amount of the compound as the free base.

Pharmaceutical Compositions

While it is possible that, for use in therapy, a therapeuticallyeffective amount of a compound disclosed herein may be administered asthe raw chemical, it is typically presented as the active ingredient ofa pharmaceutical composition or formulation. Accordingly, the disclosurefurther provides a pharmaceutical composition comprising a compounddisclosed herein. The pharmaceutical composition may further compriseone or more pharmaceutically acceptable carriers, diluents, and/orexcipients. The carrier(s), diluent(s) and/or excipient(s) must beacceptable in the sense of being compatible with the other ingredientsof the formulation and not deleterious to the recipient thereof. Inaccordance with another aspect of the disclosure there is also provideda process for the preparation of a pharmaceutical formulation includingadmixing a compound disclosed herein with one or more pharmaceuticallyacceptable carriers, diluents and/or excipients.

Pharmaceutical formulations may be presented in unit dose formscontaining a predetermined amount of active ingredient per unit dose.Such a unit may contain, for example, 0.5 mg to 1 g, preferably 1 mg to700 mg, more preferably 5 mg to 100 mg of a compound disclosed herein(as a free-base, solvate (including hydrate) or salt, in any form),depending on the condition being treated, the route of administration,and the age, weight and condition of the patient. Preferred unit dosageformulations are those containing a daily dose, weekly dose, monthlydose, a sub-dose or an appropriate fraction thereof, of an activeingredient. Furthermore, such pharmaceutical formulations may beprepared by any of the methods well known in the pharmacy art.

Pharmaceutical formulations may be adapted for administration by anyappropriate route, for example by the oral (including capsules, tablets,liquid-filled capsules, disintegrating tablets, immediate, delayed andcontrolled release tablets, oral strips, solutions, syrups, buccal andsublingual), rectal, nasal, inhalation, topical (including transdermal),vaginal or parenteral (including subcutaneous, intramuscular,intravenous or intradermal) route. Such formulations may be prepared byany method known in the art of pharmacy, for example by bringing intoassociation the active ingredient with the carrier(s), excipient(s) ordiluent. Generally, the carrier, excipient or diluent employed in thepharmaceutical formulation is “non-toxic,” meaning that it/they is/aredeemed safe for consumption in the amount delivered in thepharmaceutical composition, and “inert” meaning that it/they does/do notappreciably react with or result in an undesired effect on thetherapeutic activity of the active ingredient.

Pharmaceutical formulations adapted for oral administration may bepresented as discrete units such as liquid-filled or solid capsules;immediate, delayed or controlled release tablets; powders or granules;solutions or suspensions in aqueous or non-aqueous liquids; edible foamsor whips; oil-in-water liquid emulsions, water-in-oil liquid emulsionsor oral strips, such as impregnated gel strips.

For instance, for oral administration in the form of a tablet orcapsule, the active drug component can be combined with an oralpharmaceutically acceptable carrier such as ethanol, glycerol, water andthe like. Powders are prepared by comminuting the compound to a suitablefine size and mixing with a similarly comminuted pharmaceutical carriersuch as an edible carbohydrate, as, for example, starch or mannitol.Flavoring, preservative, dispersing and coloring agent can also bepresent.

Solid capsules are made by preparing a powder mixture, as describedabove, and filling formed gelatin sheaths. Glidants and lubricants suchas colloidal silica, talc, magnesium stearate, calcium stearate or solidpolyethylene glycol can be added to the powder mixture before thefilling operation. A disintegrating or solubilizing agent such asagar-agar, calcium carbonate or sodium carbonate can also be added toimprove the availability of the medicament when the capsule is ingested.

Moreover, when desired or necessary, suitable binders, lubricants,disintegrating agents and coloring agents can also be incorporated intothe mixture. Suitable binders include starch, gelatin, natural sugarssuch as glucose or beta-lactose, corn sweeteners, natural and syntheticgums such as acacia, tragacanth or sodium alginate,carboxymethylcellulose, polyethylene glycol, waxes and the like.Lubricants used in these dosage forms include sodium oleate, sodiumstearate, magnesium stearate, sodium benzoate, sodium acetate, sodiumchloride and the like. Disintegrators include, without limitation,starch, methyl cellulose, agar, bentonite, xanthan gum and the like.Tablets are formulated, for example, by preparing a powder mixture,granulating or slugging, adding a lubricant and disintegrant andpressing into tablets. A powder mixture is prepared by mixing thecompound, suitably comminuted, with a diluent or base as describedabove, and optionally, with a binder such as carboxymethylcellulose, analginate, gelatin, or polyvinyl pyrrolidone, a solution retardant suchas paraffin, a resorption accelerator such as a quaternary salt and/oran absorption agent such as bentonite, kaolin or dicalcium phosphate.The powder mixture can be granulated by wetting with a binder such assyrup, starch paste, acadia mucilage or solutions of cellulosic orpolymeric materials and forcing through a screen. As an alternative togranulating, the powder mixture can be run through the tablet machineand the result is imperfectly formed slugs broken into granules. Thegranules can be lubricated to prevent sticking to the tablet formingdies by means of the addition of stearic acid, a stearate salt, talc ormineral oil. The lubricated mixture is then compressed into tablets. Thecompounds disclosed herein can also be combined with a free flowinginert carrier and compressed into tablets directly without going throughthe granulating or slugging steps. A clear or opaque protective coatingconsisting of a sealing coat of shellac, a coating of sugar or polymericmaterial and a polish coating of wax can be provided. Dyestuffs can beadded to these coatings to distinguish different unit dosages.

Oral fluids such as solutions, syrups and elixirs can be prepared indosage unit form so that a given quantity contains a predeterminedamount of the compound. Solutions and syrups can be prepared bydissolving the compound in a suitably flavored aqueous solution, whileelixirs are prepared through the use of a pharmaceutically acceptablealcoholic vehicle. Suspensions can be formulated by dispersing thecompound in a pharmaceutically acceptable vehicle. Solubilizers andemulsifiers such as ethoxylated isostearyl alcohols and polyoxy ethylenesorbitol ethers, preservatives, flavor additive such as peppermint oilor natural sweeteners or saccharin or other artificial sweeteners, andthe like can also be added.

Where appropriate, unit dosage formulations for oral administration canbe microencapsulated. The formulation can also be prepared to prolong orsustain the release as for example by coating or embedding particulatematerial in polymers, wax or the like.

The compounds of the disclosure can also be administered in the form ofliposome delivery systems, such as small unilamellar vesicles, largeunilamellar vesicles and multilamellar vesicles. Liposomes can be formedfrom a variety of phospholipids, such as cholesterol, stearylamine orphosphatidylcholines.

Pharmaceutical formulations adapted for topical administration in themouth include lozenges, pastilles and mouth washes.

Pharmaceutical formulations adapted for rectal administration may bepresented as suppositories or as enemas.

Pharmaceutical formulations adapted for nasal administration wherein thecarrier is a solid include a coarse powder having a particle size forexample in the range 20 to 500 microns which is administered in themanner in which snuff is taken, i.e. by rapid inhalation through thenasal passage from a container of the powder held close up to the nose.Suitable formulations wherein the carrier is a liquid, foradministration as a nasal spray or as nasal drops, include aqueous oroil solutions of the active ingredient.

Pharmaceutical formulations adapted for administration by inhalationinclude fine particle dusts or mists, which may be generated by means ofvarious types of metered dose pressurized aerosols, metered doseinhalers, dry powder inhalers, nebulizers or insufflators.

Pharmaceutical formulations adapted for vaginal administration may bepresented as pessaries, tampons, creams, gels, pastes, foams or sprayformulations.

Pharmaceutical formulations adapted for parenteral administrationinclude aqueous and non-aqueous sterile injection solutions which maycontain anti-oxidants, buffers, bacteriostats and solutes which renderthe formulation of pharmaceutically acceptable tonicity with the bloodof the intended recipient; and aqueous and non-aqueous sterilesuspensions which may include suspending agents and thickening agents.The formulations may be presented in unit-dose or multi-dose containers,for example sealed ampoules and vials, and may be stored in afreeze-dried (lyophilized) condition requiring only the addition of thesterile liquid carrier, for example water for injection, immediatelyprior to use. Extemporaneous injection solutions and suspensions may beprepared from sterile powders, granules and tablets.

It should be understood that in addition to the ingredients particularlymentioned above, the formulations may include other agents conventionalin the art having regard to the type of formulation in question, forexample, those suitable for oral administration may include flavoringagents.

Examples Crystal Structure of RJW100 Bound to LRH-1

To understand how RJW100 interacts with LRH-1 and affects receptorconformation, the x-ray crystal structure of LRH-1 LBD bound to theagonist and to a fragment of the co-activator, Tif2, to a resolution of1.85 Å was determined (FIG. 3A). Although the RJW100 used forcrystallization was a racemic mixture of two exo stereoisomers (FIG. 1),the electron density in the structure unambiguously indicates that asingle enantiomer is bound (FIG. 3B). The bound isomer has Rstereochemistry at both the 1-position (hydroxyl-substituted) and3a-position (styrene-substituted) (hereafter RR-RJW100, FIG. 1B). Theligand is bound at a single site deep in the binding pocket and is fullyengulfed within it. This binding mode is markedly different from that ofthe PL ligands, DLPC and PIP3, which extend lower in the pocket with theheadgroups protruding into the solvent (seen by superposition with PDBs4DOS and 4RWV, respectively, FIGS. 3C and D). PL ligands also increasethe pocket volume and width compared with RJW100. For example, the mouthof the pocket is ˜3 Å wider and nearly 40% larger in volume when DLPC isbound versus RJW100 (FIG. 3E). This effect appears to be mainly due to ashift of H6, which swings away from the mouth of the pocket in the DLPCstructure by ˜3 Å(FIG. 3E). The direction and magnitude of the H6movement are similar in other LRH-1-PL structures; comparison of fourpublished human LRH-1-PL structures shows an average H6 shift of 3.0±0.2Å relative to LRH-1 in the apo-state or when synthetic ligands arebound. Although these structures exhibit diverse types of crystalpacking, the movement of H6 appears to be related to whether the ligandis a PL or small molecule and not to crystal form or packing contacts.It likely occurs to avoid stearic clashes with the PL headgroup.Notably, the H6/0-sheet region has been recently identified as a sitethrough which PL ligands allosterically communicate with the AFS tomodulate LRH-1 activity. The fact that the synthetic agonists do notdisplace H6 relative to the apo-receptor suggests that they utilize adifferent mechanism for receptor activation.

Repositioning of RJW100 Compared with a Closely Related SyntheticAgonist

Perhaps the most striking observation from structure comes fromcomparison of RJW100 with GSK8470-bound LRH-1 (PBD 3PLZ). Overall,protein conformation is highly similar; the largest movement occurs inthe bottom of H3, which moves in the direction of H6 (by 2 Å in theRR-RJW100 structure and by 4 Ain the GSK8470 structure relative toapo-LRH-1). However, there is a substantial difference in thepositioning of these agonists within the binding pocket. AlthoughGSK8470 and RR-RJW100 bind in the same vicinity, they are rotated nearly180° from one another. The bicyclic rings at the cores of each moleculeare perpendicular to each other, causing the tails to be pointed inopposite directions (FIG. 4A). Notably, the rationale for adding ahydroxyl group in the 1-position on this scaffold was to promote aninteraction with a “polar patch,” consisting of residues Arg-393 andis-390 in an otherwise hydrophobic pocket. This interaction waspredicted based on the position of the ligand in the LRH-1-GSK8470structure; however, the actual position of RR-RJW100 in the pocketplaces the hydroxyl group over 6 Å away from these residues (FIG. 4B).Such a radically different binding mode for closely related moleculeswas unexpected, and a propensity to rotate within the pocket maycontribute to difficulties improving agonist activity by modification ofthe GSK8470 scaffold.

Discovery of a LRH-1 Interaction Mediated by the RJW100 Hydroxyl Group

Protein-ligand interactions made by GSK8470 and the RJW100 stereoisomerswere examined to gain insight into factors influencing theligand-binding mode. A close view of the LRH-1-binding pocket revealsthat RR-RJW100 makes several hydrophobic contacts, many of which arealso made by GSK8470 (shown in, FIG. 5A). Additionally, RR-RJW100 makesseveral unique contacts (shown in FIG. 5A). Many of these uniquecontacts are also hydrophobic; however, the RR-RJW100 hydroxyl groupforms an indirect polar contact with residue Thr-352 via a watermolecule. A portion of the electron density map is shown in FIG. 5A toemphasize the strong evidence for this interaction. SR-RJW100 alsointeracts with Thr-352 through the same water molecule, despite thediffering conformations of the hydroxyl group (FIG. 5B). The position ofthe SR-RJW100 hydroxyl group also permits a second water-mediatedhydrogen-bonding interaction with the backbone nitrogen of residueVal-406 (FIG. 5B).

Although the interaction with Thr-352 is indirect, the water moleculeinvolved is part of a network of waters found in every LRH-1 crystalstructure in the same location. Additional support for modeling water atthese sites in the pocket comes from B-factors of ligating atoms. Thus,this water network appears to be a conserved feature of the bindingpocket and may play a role in receptor function or stability. To testthe hypothesis that the OH-water-Thr-352 interaction was influencingligand positioning, the stability of this bond was analyzed usingmolecular dynamics simulations (MDS). Throughout each simulation (200ns), the four conserved networked water molecules remained in the samepositions (if a particular water molecule occasionally left, it wasimmediately replaced with another in the same location). Residue Thr-352maintained a hydrogen bond with the water molecule for 100% of eachsimulation, regardless of which ligand was bound. Additionally, bothRR-RJW100 and SR-RJW100 maintained hydrogen bonding with the watermolecule for the majority of the simulations (53.7% of the time forRR-RJW100 and 64.4% of the time for SR-RJW100). When residue Thr-352 wasmutated to valine in MDS, the time spent interacting with theThr-352-coordinated water molecule was drastically reduced (22.9 and0.5% when RR-RJW100 and SR-RJW100 were bound, respectively),demonstrating that this mutation likely disrupts this water-mediatedinteraction made by these ligands.

Differences in π-π Stacking with Residue His-390 among LRH-1 Agonists

The π-π-stacking of GSK8470 with residue His-390 has been described asimportant for activation of LRH-1 by synthetic compounds. The RJW100diastereomers also engage in π-π-stacking with His-390, but with somekey differences. The π-π-stacking is face-to-face for GSK8470 andedge-to-face for the RJW100 isomers. Additionally, by virtue of the verydifferent orientations in the binding pocket, the agonists do not useanalogous phenyl rings for π-π-stacking; GSK8470 uses the aniline group,whereas the RJW100 isomers use the adjacent phenyl substituent.Moreover, MDS demonstrate ligand-dependent differences in the stabilityof this interaction.

Role of Thr-352 and his-390 in LRH-1 Activation by Synthetic Agonists

The importance of the Thr-352 and His-390 interactions for binding andactivation of LRH-1 by the agonists was investigated using mutagenesis.Binding and stabilization of LRH-1 were detected using DSF. Although theT352V mutation (designed to remove the water-mediated hydrogen bond withbound ligands) had little effect on the overall thermostability ofDLPC-bound LRH-1, it completely abrogated the stabilizing effect ofRR-RJW100 and SR-RJW100. Likewise, disrupting this interaction by usingan RJW100 analog lacking the hydroxyl group prevented the positive Tmshift in wild-type (WT) LRH-1 (FIG. 6A). GSK8470 did not affect themelting profile of LRH-1 in WT or T352V protein, supporting the notionthat the hydroxyl group is important for stabilizing the protein-ligandcomplex.

The Thr-352 interaction was also found to be important for LRH-1activation by small molecule agonists. The Compound lacking the hydroxylgroup and unable to make this interaction, was an extremely poor LRH-1activator in luciferase reporter assays. The endo-RJW100 was also a weakagonist, although statistically significant activation was achieved atthe highest dose with WT LRH-1 (˜1.4-fold over DMSO, FIG. 6D). RR-RJW100and GSK8470 were equally effective toward WT LRH-1, and both increasedactivity by ˜2.5-fold compared with DMSO at the highest dose, and bothhad EC₅₀ values of around 4 m (FIGS. 6B and 6C). Notably, the T352Vmutation greatly reduced the ability of RR-RJW100 to activate LRH-1compared with WT protein, while not significantly affecting baselineactivity. Unexpectedly, this mutation similarly attenuated activation byGSK8470, perhaps suggesting a broader role for this residue (or perhapsfor the water network it coordinates) in ligand-mediated activation.Indeed, introduction of a T352V mutation to GSK8470-bound LRH-1 in MDSdisrupts the water network, causing complete displacement of the watermolecule typically coordinated by Thr-352. The T352V mutation alsosignificantly reduces the amount of time GSK8470 spends π-π-stackingwith His-390 (25.7% versus 89.5% of the simulation). Both thedestabilization of the water network and the disruption of stableHis-390 π-π-stacking by the T352V mutation could contribute to theobserved loss of activity for GSK8470 in the context of this mutation.

Although the T352V mutation resulted in a loss of activity for bothRR-RJW100 and GSK8470, mutating His-390 to alanine had a differenteffect on LRH-1 activation depending on the agonist involved. GSK8470was completely unable to activate H390A-LRH-1, but this mutation hadlittle to no effect on RR-RJW100-mediated activation. This differentialreliance on His-390 for activation is consistent with the observationthat GSK8470 interacts with His-390 more stably than RR-RJW100 in MDS.This also provides evidence that RR-RJW100 utilizes a differentmechanism of action than GSK8470 for LRH-1 activation.

Synthesis of Hexahydropentalene Derivatives

The general strategy for preparing hexahydropentalene derivatives isillustrated in FIG. 1 using procedures set out in, or as appropriatelymodified from, Whitby et al. (2011). Small molecule agonists of theorphan nuclear receptors steroidogenic factor-1 (SF-1, NR5A1) and liverreceptor homologue-1 (LRH-1, NR5A2). Journal of Medicinal Chemistry, 54,2266-2281.

Synthetic Scheme for Enyne (4)

5-phenylpent-4-yn-1-ol, 99%

To an oven-dried round bottom flask was addedbis(triphenylphosphine)palladium dichloride (0.03 equiv.) and copperiodide (0.06 equiv.). Triethylamine was added to make a 1.0 M solutionbefore the addition of iodobenzene (1.0 equiv.) The resulting yellowmixture was sparged by bubbling the solution with nitrogen for a periodof 30 minutes, at which point 4-pentyl-1-ol (1.2 equiv.) was addedportionwise and the sparging needle was replaced with anitrogen inlet.The solution rapidly darkened and formed a slurry, and was heated at 60C for 2 hours, at which point the reaction was complete by TLC. Theresulting black solution was cooled and ether was added to precipitate ablack solid. The resulting slurry was filtered over a plug of celite andeluted with ether. The filtrate was concentrated in vacuo to afford areddish-brown oil, which was then purified on silica in 30% EtOAc/hex toafford a red oil.

5-phenylpent-4-ynal, 83%

To an oven-dried 3-neck flask under nitrogen, oxalyl chloride (1.1equiv.) in DCM (0.1M) was cooled to −78 C in a dry ice/acetone bath.Dimethylsulfoxide (DMSO) (1.3 equiv.) was added dropwise as a solutionin DCM. The solution was allowed to stir approximately 10 minutes untilbubbling ceased. The required alcohol was then added dropwise as asolution in DCM. The reaction mixture was stirred and maintained at −78C for 1 hour. Triethylamine (2.5 equiv.) was then added at −78 C andallowed to warm to room temperature. The reaction was quenched withsaturated aqueous ammonium chloride and extracted with ethyl acetate.The combined organics were washed with brine, dried with MgSO₄ andconcentrated in vacuo. The resulting residue was purified on silica in10-20% EtOAc/Hexanes to afford a clear, yellow oil.

7-phenylhept-1-en-6-yn-3-ol, 81%

To an oven-dried 3-neck flask under nitrogen was added aldehyde, (1.0equiv.) as a solution in dry THF. The solution as cooled to −78 C andvinylmagnesium bromide (1.0 M, 1.5 equiv.) was added. The reactionmixture was stirred and allowed to warm to room temperature over 5 h.The reaction was quenched with ammonium chloride, poured onto water, andextracted with ethyl acetate. The combined organic layers were washedwith brine, dried with MgSO₄ and concentrated in vacuo. The resultingoil was purified on silica in 5-10% EtOAc/Hexanes to afford a clear,colorless oil.

(5-(methoxymethoxy)hept-6-en-1-ynl)benzene (4), 68%

Allylic alcohol (1.0 equiv.) was dissolved in DCM and cooled to 0 C.Diisopropylethyl amine (1.25 equiv.) was added, followed bychloro(methoxy)methane (1.5 equiv.). The reaction mixture was stirredover 18 hours, and then poured over water and extracted with DCM. Thecombined organic layers were washed with 1M HCl, then brine, then driedover MgSO₄ and concentrated in vacuo. The resulting oil was purified in5% EtOAc/Hexanes to afford a clear, colorless oil.

Synthesis of Dibromo Compounds

General Procedure for the Formation of TBDPS-Protected Diols

To a round bottom flask open to air the required diol (2.0 equiv.) wasdissolved in THE to make a 0.1M solution. Imidazole (1.0 equiv.) wasadded, followed by tert-butyl diphenyl silyl chloride (TBDPSCl) (1.0equiv.). The resulting solution was allowed to stir for 18 h, upon whicha solid white precipitate formed. The solution was filtered and thefiltrate concentrated in vacuo. The resulting material was subjected tosilica gel chromatography in 5-150% EtOAc/Hexanes to afford a clear,colorless oil.

n IUPAC Name Yield (%) 1 5-((tert-butyldiphenylsilyl)oxy)pentan-1-ol 5a88 2 6-((tert-butyldiphenylsilyl)oxy)hexan-1-ol 6a 40 37-((tert-butyldiphenylsilyl)oxy)heptan-1-ol 7a 38 48-((tert-butyldiphenylsilyl)oxy)octan-1-ol 8a 74 59-((tert-butyldiphenylsilyl)oxy)nonan-1-ol 9a 82 610-((tert-butyldiphenylsilyl)oxy)decan-1-ol 10a 81 711-((tert-butyldiphenylsilyl)oxy)undecan-1-ol 11a 82 812-((tert-butyldiphenylsilyl)oxy)dodecan-1-ol 12a 35

General Procedure for the Formation of TBDPS Protected Aldehydes

To around bottom flask open to air the required TBDPS-monoprotected diol(1a-8a) was dissolved in dichloromethane to form a 0.1M solution.Trichloroisocyanuric acid (1.0 equiv.) was added to the reactionmixture, followed by (2,2,6,6,-Tetramethyl-piperidin-1-yl)oxyl (TEMPO)(0.01 equiv.). The reaction was monitored by TLC until consumption ofthe starting diol was consumed (less than 10 minutes). The resultingsolution was poured over water and quenched with saturated sodiumbicarbonate. The aqueous layer was extracted with dichloromethane threetimes. The combined organic layers were then washed with 1M HCl andbrine, after which the organic layer was dried with MgSO₄ andconcentrated in vacuo. The resulting oil was subjected to silica gelchromatography in 20% EtOAc/Hexanes to afford a clear, colorless oil.

n IUPAC Name Yield (%) 1 5-((tert-butyldiphenylsilyl)oxy)pentanal (5b)90 2 6-((tert-butyldiphenylsilyl)oxy)hexanal (6b) 82 37-((tert-butyldiphenylsilyl)oxy)heptanal (7b) 93 48-((tert-butyldiphenylsilyl)oxy)octanal (8b) 57 5 9-((tert-butyldiphenylsilyl)oxy)nonanal (9b) 63 6 10-((tert-butyldiphenyl silyl)oxy)decanal(10b) 88 7 11-((tert-butyldiphenylsilyl)oxy)undecanal (11b) 60 812-((tert-butyldiphenylsilyl)oxy)dodecanal (12b) 95General Procedure for the Formation of Terminal Gem-Dibromo Compounds1c-8c

Triphenylphosphite (1.1 equiv) was added to an oven-dried three-neckflask under nitrogen, dissolved in dichloromethane (DCM), and cooled to−78 C in a dry ice/acetone bath. Bromine (1.1 equiv) was addedportionwise. Triethylamine (1.1 equiv.) was added dropwise and thesolution was allowed to stir for 5 minutes. The required aldehyde(1b-8b) was added as a solution in dichloromethane and the mixture wasallowed to stir for hours. Upon completion, the reaction mixture wasfiltered over a plug of silica (eluted with ethyl acetate) and thefiltrate was concentrated in vacuo. The resulting oil was subjected to ashort plug of silica and eluted with 100% hexanes to afford a clear,colorless oil.

IUPAC Name Yield (%) tert-butyl((5,5-dibromopentyl)oxy)diphenylsilane(5c) 73 tert-butyl((6,6-dibromohexyl)oxy)diphenylsilane (6c) 74tert-butyl((7,7-dibromoheptyl)oxy)diphenylsilane (7c) 45tert-butyl((8,8-dibromooctyl)oxy)diphenylsilane (8c) 81tert-butyl((9,9-dibromononyl)oxy)diphenylsilane (9c) 89tert-butyl((10,10-dibromodecyl)oxy)diphenylsilane (10c) 55tert-butyl((11,11-dibromoundecyl)oxy)diphenylsilane (11c) 63tert-butyl((12,12-dibromododecyl)oxy)diphenylsilane (12c) 65

Bis-Protected 5,5-Bicyclic Compounds

General Procedure for MOM-Protected 2°, Deprotected 1° Alcohols (5d-12d)

Bis(cyclopentadienyl)zirconium(IV) dichloride (zirconecene dichloride)(1.2 equiv.) was dried by azeotroping away latent water with benzenefour times before being placed under nitrogen, dissolved in dry,degassed tetrahydrofuran (THF) and cooled to −78° C. in a dryice/acetone bath. The resulting solution of zirconecene dichloride wastreated with nBuLi (1.6M in hex, 2.4 equiv.) to form a clear, lightyellow solution and allowed to stir. After approximately 30 minutes,azeotroped (5-(methoxymethoxy)hept-6-en-1-yn-1-yl)benzene (4) (1.0equiv.) in dry, degassed THF was added portionwise to afford apink-orange solution, and the reaction mixture was held at −78° C. for30 minutes before allowing to warm to room temperature and stirred over2.5 hours. The reaction mixture was then re-cooled to −78° C. and therequired azeotroped required 1,1-dibromoalkane protected alcohol(5c-12c) (1.1 equiv.) were added in dry, degassed THF. Freshly preparedlithium diisopropylamine (LDA, 1.0 M, 1.1 equiv.) was added at −78° C.and stirred for 15 minutes. Freshly prepared lithium phenylacetylide(3.6 equiv.) was added to the reaction mixture dropwise in dry, degassedTHF. The resulting dark reddish brown solution was stirred at −78° C.for 1.5 hours. The reaction was then quenched with methanol andsaturated aqueous sodium bicarbonate and allowed to warm to roomtemperature to form alight yellow slurry. The slurry was poured overwater and extracted with ethyl acetate four times. The combined organiclayers were washed with brine, dried with MgSO₄, and concentrated invacuo. The resulting colored yellow oil was roughly purified on a plugof silica and eluted with 20% EtOAc/Hexanes to afford an oil which is amixture of quenched phenylacetylide, desired bis-protected [3.3.0]bicyclic compounds (and in some cases protonolysis byproduct), which wascarried on without further purification. The crude oil was thendissolved in a round bottom flask charged with a stir bar and open toair. TBAF (approx. 2.0 equiv. of enyne starting material) was added. Thesolution rapidly darkened and was allowed to stir at room temperaturefor 16 h. After reaction completion, the reaction mixture wasconcentrated and directly subjected to purification by silica gelchromatography to afford the desired cyclized, free primary alcoholproducts 5d-12d.

Identifier Compound Name of Major Product % Yield (2 steps)  5d4-(6-exo-(methoxymethoxy)-3-phenyl-3a-(1- 40phenylvinyl)-1,3a,4,5,6,6a-hexahydropentalen-2-yl)butan-1-ol  6d5-(6-exo-(methoxymethoxy)-3-phenyl-3a-(1- 62phenylvinyl)-1,3a,4,5,6,6a-hexahydropentalen-2-yl)pentan-1-ol  7d6-(6-exo-(methoxymethoxy)-3-phenyl-3a-(1- 80phenylvinyl)-1,3a,4,5,6,6a-hexahydropentalen-2-yl)hexan-1-ol  8d7-(6-exo-(methoxymethoxy)-3-phenyl-3a-(1- 42phenylvinyl)-1,3a,4,5,6,6a-hexahydropentalen-2-yl)heptan-1-ol  9d8-(6-exo-(methoxymethoxy)-3-phenyl-3a-(1- 72phenylvinyl)-1,3a,4,5,6,6a-hexahydropentalen-2-yl)octan-1-ol 10d9-(6-exo-(methoxymethoxy)-3-phenyl-3a-(1- 53phenylvinyl)-1,3a,4,5,6,6a-hexahydropentalen-2-yl)nonan-1-ol 11d10-(6-exo-(methoxymethoxy)-3-phenyl-3a-(1- 60phenylvinyl)-1,3a,4,5,6,6a-hexahydropentalen-2-yl)decan-1-ol 12d11-(6-exo-(methoxymethoxy)-3-phenyl-3a-(1- 70phenylvinyl)-1,3a,4,5,6,6a-hexahydropentalen-2-yl)undecan-1-olGeneral Procedure for Diols (5e-12e)

To a solution of required cyclized primary alcohols (5d-12d) inacetonitrile was added concentrated HCl in excess (5-20 equiv.). Thesolution was stirred for 18 h or until the reaction was completed byTLC. The resulting solution as concentrated in vacuo and subjected topreparatory HPLC to isolate the desired product as the major exodiastereomer.

Identi- fier Compound Name  5eExo-5-(4-hydroxybutyl)-4-phenyl-3a-(1-phenylvinyl)-1,2,3,3a,6,6a-hexahydropentalen-1-ol  6eExo-5-(5-hydroxypentyl)-4-phenyl-3a-(1-phenylvinyl)-1,2,3,3a,6,6a-hexahydropentalen-1-ol  7eExo-5-(6-hydroxyhexyl)-4-phenyl-3a-(1-phenylvinyl)-1,2,3,3a,6,6a-hexahydropentalen-1-ol  8eExo-5-(7-hydroxyheptyl)-4-phenyl-3a-(1-phenylvinyl)-1,2,3,3a,6,6a-hexahydropentalen-1-ol  9eExo-5-(8-hydroxyoctyl)-4-phenyl-3a-(1-phenylvinyl)-1,2,3,3a,6,6a-hexahydropentalen-1-ol 10eExo-5-(9-hydroxynonyl)-4-phenyl-3a-(1-phenylvinyl)-1,2,3,3a,6,6a-hexahydropentalen-1-ol 11eExo-5-(10-hydroxydecyl)-4-phenyl-3a-(1-phenylvinyl)-1,2,3,3a,6,6a-hexahydropentalen-1-ol 12eExo-5-(11-hydroxyundecyl)-4-phenyl-3a-(1-phenylvinyl)-1,2,3,3a,6,6a-hexahydropentalen-1-ol

Characterization Data for Diols

5e:

¹H NMR (600 MHz, Chloroform-d) δ 7.33-7.24 (m, 6H), 7.23 (s, 2H),7.20-7.16 (m, 2H), 5.05 (d, J=1.4 Hz, 1H), 4.97 (d, J=1.5 Hz, 1H), 3.93(s, 1H), 3.55 (t, J=6.2 Hz, 2H), 2.36 (dd, J=16.9, 9.3 Hz, 1H), 2.29 (d,J=6.9 Hz, 1H), 2.10-2.02 (m, 4H), 1.74-1.61 (m, 3H), 1.50-1.37 (m, 4H).¹³C NMR (500 MHz, Chloroform-d) 6154.60, 144.10, 140.77, 139.42, 137.31,129.62, 127.71, 127.67, 127.63, 126.61, 116.56, 114.84, 81.80, 69.29,62.39, 55.65, 40.13, 33.94, 32.65, 32.01, 29.44, 24.03, 22.45, 10.57.HRMS calcd for C26H31O2 [M+H]+: 375.23186, found 375.23145 IR (cm-1):3344 (b), 3079 (w), 2937, 2191 (w), 1737, 1669, 1597, 1490, 1440, 1378,1349, 1229, 1217, 1070, 1028.

6e:

¹H NMR (500 MHz, Chloroform-d) δ 7.43-7.28 (m, 7H), 7.28-7.19 (m, 3H),5.09 (d, J=1.4 Hz, 1H), 5.01 (d, J=1.4 Hz, 1H), 3.97 (s, 1H), 3.61 (t,J=6.6 Hz, 2H), 2.38 (dd, J=16.8, 9.4 Hz, 1H), 2.31 (d, J=9.3, 1.5 Hz,1H), 2.16-2.03 (m, 4H), 1.77-1.65 (m, 3H), 1.56-1.48 (m, 3H), 1.43-1.25(m, 3H). ¹³C NMR (500 MHz, Chloroform-d) δ 154.50, 144.12, 140.79,139.31, 137.27, 131.51, 129.64, 128.19, 127.72, 127.64, 126.66, 126.64,115.04, 82.03, 69.32, 62.85, 61.85, 55.76, 40.25, 33.98, 32.51, 32.05,31.33, 29.65, 27.62, 25.75, 15.98. HRMS calcd for C27H33O2 [M+H]+:389.24751, found 398.24762 IR (cm-1): 3332 (b), 3079 (w), 2933, 1490,1441, 1342, 1191, 1071, 1035.

7e:

¹H NMR (600 MHz, Chloroform-d) δ 7.36-7.25 (m, 6H), 7.24-7.15 (m, 4H),5.05 (s, 1H), 4.97 (s, 1H), 3.93 (s, 1H), 3.58 (t, J=6.6 Hz, 2H), 2.35(dd, J=16.9, 9.4 Hz, 1H), 2.28 (d, J=9.4 Hz, 1H), 2.11-1.97 (m, 4H),1.75-1.62 (m, 3H), 1.53-1.46 (m, 2H), 1.38-1.15 (m, 6H). ¹³C NMR (600MHz, Chloroform-d) δ 154.52, 144.14, 140.94, 139.23, 137.33, 129.68,127.70, 127.62, 126.66, 126.61, 115.04, 109.98, 82.03, 69.33, 62.96,55.76, 40.25, 33.97, 32.63, 32.08, 29.57, 29.38, 27.73, 25.46, HRMScalcd for C28H35O2 [M+H]+: 403.26316, found 403.26338 IR (cm-1) 3330(b), 3051 (w), 2929, 2855, 1490, 1440, 1340.74, 1262.75, 1191, 1071,1028.

8e:

¹H NMR (600 MHz, Chloroform-d) δ 7.35-7.25 (m, 5H), 7.24-7.15 (m, 5H),5.05 (d, J=1.4 Hz, 1H), 4.97 (d, J=1.4 Hz, 1H), 3.93 (s, 1H), 3.60 (t,J=6.7 Hz, 2H), 2.35 (dd, J=16.9, 9.3 Hz, 1H), 2.28 (d, J=9.4 Hz, 1H),2.09-1.98 (m, 5H), 1.73-1.61 (m, 3H), 1.36-1.19 (m, 10H). ¹³C NMR (600MHz, Chloroform-d) δ 154.53, 144.14, 141.03, 139.16, 137.33, 129.67,127.70, 127.61, 126.64, 126.59, 115.02, 82.03, 69.31, 63.00, 58.47,55.76, 50.89, 40.22, 33.96, 32.71, 32.07, 29.63, 29.58, 29.16, 27.71,25.61, 18.42. HRMS calcd for C29H36O2Cl [M+Cl]−: 451.24093, found451.24179 IR (cm-1): 3347 (b), 3079 (w), 3052 (w), 2929, 2855, 1491,1441, 1342, 1261, 1192, 1028.

9e:

¹H NMR (600 MHz, Chloroform-d) δ 7.35-7.25 (m, 6H), 7.24-7.16 (m, 4H),5.05 (d, J=1.4 Hz, 1H), 4.97 (d, J=1.4 Hz, 1H), 3.93 (s, 1H), 3.61 (t,J=6.7 Hz, 2H), 2.35 (dd, J=16.9, 9.3 Hz, 1H), 2.28 (d, J=9.2 Hz, 1H),2.10-2.01 (m, 2H), 2.00-1.97 (m, 2H), 1.74-1.61 (m, 3H), 1.35-1.16 (m,13H). HRMS calcd for C30H38O2Cl [M+Cl]−: 465.25658, found 465.25703 IR(cm-1): 3344 (b), 3079 (w), 3052 (w), 2927, 2854, 1491, 1440, 1343,1261, 1192, 1071, 1029.

10e:

¹H NMR (600 MHz, Chloroform-d) δ 7.39-7.26 (m, 5H), 7.24-7.14 (m, 5H),5.05 (d, J=1.4 Hz, 1H), 4.97 (d, J=1.4 Hz, 1H), 3.93 (s, 1H), 3.62 (t,J=9.4 Hz, 2H), 2.34 (dd, J=16.7, 9.4 Hz, 1H), 2.27 (d, J=9.3 Hz, 1H),2.12-2.03 (m, 2H), 2.01-1.96 (m, 1H), 1.74-1.58 (m, 3H), 1.34-1.17 (m,18H). ¹³C NMR (600 MHz, Chloroform-d) δ 154.56, 144.15, 141.13, 139.08,137.35, 129.68, 127.71, 127.70, 127.59, 126.64, 126.57, 115.01, 82.05,69.32, 63.06, 55.77, 40.23, 33.97, 32.76, 32.08, 29.66, 29.61, 29.46,29.36, 29.31, 27.78, 25.67. HRMS calcd for C31H40O2Cl [M+Cl]−:479.27223, 479.27260 IR (cm-1): 3343 (b), 3079 (w), 3052 (w), 2928,2854, 1670.41, 1598, 1491, 1440, 1344, 1192, 1071, 1055, 1029.

11e:

¹H NMR (600 MHz, Chloroform-d) δ 7.35-7.25 (m, 5H), 7.24-7.16 (m, 5H),5.05 (d, J=1.4 Hz, 1H), 4.97 (d, J=1.4 Hz, 1H), 3.93 (s, 1H), 3.61 (t,J=6.7 Hz, 2H), 2.34 (dd, J=16.9, 9.4 Hz, 1H), 2.27 (d, J=9.2 Hz, 1H),2.11-2.00 (m, 3H), 1.73-1.62 (m, 3H), 1.54 (dq, J=8.2, 6.7 Hz, 2H),1.34-1.17 (m, 17H). LRMS [APCI] calcd for C32H41O2 [M−H]−: 457.3, found457.2

12e:

¹H NMR (600 MHz, Chloroform-d) δ 7.34-7.25 (m, 4H), 7.24-7.16 (m, 5H),5.05 (d, J=1.4 Hz, 1H), 4.96 (d, J=1.3 Hz, 1H), 3.93 (s, 1H), 3.61 (t,J=6.7 Hz, 2H), 2.33 (dd, J=17.2, 9.8 Hz, 1H), 2.26 (d, J=9.3 Hz, 1H),2.09-1.99 (m, 1H), 2.02-1.99 (m, 5H), 1.72-1.61 (m, 3H), 1.57-1.51 (m,2H), 1.40-1.11 (m, OH). LRMS [APCI] calcd for C33H43O2 [M−H]−: 471.3,found 471.0

Synthetic Scheme for [3.3.0] Bicyclic Carboxylic Acids

General Procedure for Carboxylic Acids (5f-12f)

A solution of tetrapropylammoniumperruthate (TPAP) (0.1 equiv.) andn-methylmorpholine oxide (NMO) (10.0 equiv.) in acetonitrile was madeand added to the required cyclized primary alcohol (5d-12d) (1.0 equiv.)in a scintillation vial open to air. Reagent grade water (10.0 equiv.)was added and the solution was allowed to stir for 3 hours at roomtemperature, or until the reaction was complete by TLC. The resultingblack solution was concentrated in vacuo and passed through a short plugof silica and eluted with 50% EtOAc/Hexanes with 0.1% Acetic Acid addedto afford the protected carboxylic acid intermediates.

The resulting clear, colorless oil was subsequently taken up inacetonitrile, and concentrated HCl (5-10 equiv.) was added. The solutionwas allowed to stir for 30 minutes or until complete by TLC. Theresulting solution was concentrated in vacuo and subjected topreparatory HPLC to isolate the desired product as the major exodiastereomer.

Yield Identi- (2 fier Compound Name steps)  5f4-(6-exo-hydroxy-3-phenyl-3a-(1-phenylvinyl)- 531,3a,4,5,6,6a-hexahydropentalen-2-yl)butanoic acid  6f5-(6-exo-hydroxy-3-phenyl-3a-(1-phenylvinyl)- 781,3a,4,5,6,6a-hexahydropentalen-2-yl)pentanoic acid  7f6-(6-exo-hydroxy-3-phenyl-3a-(1-phenylvinyl)- 741,3a,4,5,6,6a-hexahydropentalen-2-yl)hexanoic acid  8f7-(6-exo-hydroxy-3-phenyl-3a-(1-phenylvinyl)- 981,3a,4,5,6,6a-hexahydropentalen-2-yl)heptanoic acid  9f8-(6-exo-hydroxy-3-phenyl-3a-(1-phenylvinyl)- 951,3a,4,5,6,6a-hexahydropentalen-2-yl)octanoic acid 10f9-(6-exo-hydroxy-3-phenyl-3a-(1-phenylvinyl)- 631,3a,4,5,6,6a-hexahydropentalen-2-yl)nonanoic acid 11f10-(6-exo-hydroxy-3-phenyl-3a-(1-phenylvinyl)- 641,3a,4,5,6,6a-hexahydropentalen-2-yl)decanoic acid 12f11-(6-exo-hydroxy-3-phenyl-3a-(1-phenylvinyl)- 661,3a,4,5,6,6a-hexahydropentalen-2-yl)undecanoic acid

Characterization Data for Carboxylic Acids

5f:

¹H NMR (500 MHz, CDCl₃) δ7.38-7.16 (m, 10H), 5.08 (d, J=1.4 Hz, 1H),5.00 (d, J=1.4 Hz, 1H), 3.97 (s, 1H), 2.42-2.36 (m, 1H), 2.37-2.31 (m,2H), 2.30-2.23 (m, 2H), 2.14-2.06 (m, 4H), 1.75-1.65 (m, 5H). LRMS (ESI,APCI) m/z: calc'd for C26H27O3 [M−H]− 387.2, found 387.1 HRMS calcd forC26H27O3 [M−H]−: 387.19657, found 387.19687

6f:

¹H NMR (600 MHz, CDCl₃) δ 7.56-6.96 (m, 10H), 5.05 (d, J=1.4 Hz, 1H),4.97 (d, J=1.4 Hz, 1H), 3.93 (s, 1H), 2.38-2.23 (m, 4H), 2.13-1.97 (m,5H), 1.74-1.62 (m, 3H), 1.53 (p, J=7.4 Hz, 2H), 1.44-1.32 (m, 2H).

¹³C NMR (126 MHz, CDCl3) δ 177.81, 154.47, 144.11, 140.26, 139.83,137.17, 129.65, 127.75, 127.73, 126.75, 126.71, 115.11, 82.02, 69.37,55.77, 40.12, 34.00, 33.46, 32.09, 29.29, 27.19, 24.61. LRMS (ESI, APCI)m/z: calc'd for C27H30O3 [M−H]− 401.2, found 401.5. Calc'd for C27H29O2[M−OH]+ 385.2, found 385.2 HRMS calcd for C27H31O3 [M+H]+: 403.22677,found 403.22661. IR (cm-1): 3386 (b), 3079 (w), 3052 (2), 2940, 1707(s), 1491, 1440, 1410, 1342, 1236, 1192, 1071, 1029.

7f:

¹H NMR (600 MHz, CDCl3) δ 7.49-7.05 (m, 10H), 5.05 (d, J=1.5 Hz, 1H),4.97 (d, J=1.4 Hz, 1H), 3.93 (s, 1H), 2.36-2.21 (m, 4H), 2.10-1.97 (m,5H), 1.73-1.61 (m, 3H), 1.59-1.51 (m, 2H), 1.34 (p, J=7.6 Hz, 2H),1.28-1.20 (m, 2H). LRMS (ESI, APCI) m/z: calc'd for C28H31O3 [M−H]−415.2, found 414.9. Calc'd for C28H31O2 [M−OH]+399.2, found 399.2 HRMScalcd for C28H32O3Na [M+Na]+: 439.22353, found 439.22370 IR (cm-1): 3386(b), 3079 (w), 2934, 2856, 1708 (s), 1491, 1440, 1407, 1342, 1230, 1191,1072, 1028.

8f:

¹H NMR (600 MHz, CDCl3) δ 7.37-7.14 (m, 9H), 5.05 (d, J=1.5 Hz, 1H),4.97 (d, J=1.4 Hz, 1H), 3.93 (s, 1H), 2.38-2.25 (m, 4H), 2.11-1.97 (m,4H), 1.76-1.47 (m, 4H), 1.33 (p, J=7.6 Hz, 2H), 1.29-1.14 (m, 6H). LRMS(ESI, APCI) m/z: calc'd for C29H33O3 [M−H]− 429.3, found 429.3. Calc'dfor C29H33O2 [M−OH]+413.2, found 413.28 HRMS calcd for C29H33O3 [M−H]−:429.24352, found 429.2378

9f:

¹H NMR (600 MHz, CDCl3) δ 7.81-7.13 (m, 10H), 5.05 (s, 1H), 4.97 (s,1H), 3.93 (s, 1H), 2.37-2.22 (m, 5H), 2.09-1.94 (m, 5H), 1.79-1.58 (m,6H), 1.38-1.16 (m, 5H). LRMS (ESI, APCI) m/z: calc'd for C30H35O3 [M−H]−443.3, found 443.2. Calc'd for C30H35O2 [M−OH]+ 427.3, found 427.4 HRMScalcd for C30H35O3 [M−H]−: 443.25917, found 443.25927

10f:

¹H NMR (500 MHz, CDCl3) δ 7.41-7.15 (m, 10H), 5.07 (d, J=1.4 Hz, 1H),4.99 (d, J=1.4 Hz, 1H), 3.95 (s, 1H), 2.39-2.27 (m, 5H), 2.12-1.97 (m,5H), 1.76-1.56 (m, 6H), 1.40-1.15 (m, 7H). 13C NMR (126 MHz, CDCl3) δ179.49, 154.55, 144.14, 141.10, 139.16, 137.35, 129.69, 127.74, 127.71,127.63, 126.67, 126.61, 115.00, 82.09, 69.33, 55.71, 40.23, 33.95,32.08, 29.64, 29.53, 29.16, 29.10, 29.00, 24.65. LRMS (ESI, APCI) m/z:calc'd for C31H37O3 [M−H]− 457.3, found 457.3. Calc'd for C31H37O2[M−OH]+441.3, found 440.8 HRMS (ESI) m/z: calc'd for for C31H37O3 [M−H]−457.27482, found 457.27487 FT-IR (neat): 3361, 3079, 3052, 3019, 2928,2854, 1708, 1598, 1491, 1441, 1410, 1340, 1278, 1241, 1192, 1073, 1029,903, 766, 703 cm-1.

11f:

¹H NMR (500 MHz, CDCl3) δ 7.39-7.15 (m, 10H), 5.07 (d, J=1.6 Hz, 1H),4.99 (d, J=1.6 Hz, 1H), 3.95 (s, 1H), 2.41-2.24 (m, 5H), 2.15-1.94 (m,5H), 1.75-1.50 (m, 6H), 1.42-1.09 (m, 9H). ¹³C NMR (126 MHz, CDCl3) δ179.44, 154.57, 144.15, 141.15, 139.11, 137.37, 129.69, 127.74, 127.71,127.62, 126.66, 126.60, 115.00, 82.10, 69.33, 55.71, 40.25, 33.98,33.94, 32.08, 29.65, 29.59, 29.30, 29.28, 29.19, 29.01, 27.78, 24.68.LRMS (ESI, APCI) m/z: calc'd for C32H39O3 [M−H]− 471.3, found 471.3.Calc'd for C32H39O2 [M−OH]+455.3, found 454.8 HRMS (ESI) m/z: calc'd forC32H39O3 [M−H]− 471.28819, found 471.29047. FT-IR (neat): 3360, 3079,3052, 3019, 2926, 2854, 1708, 1598, 1491, 1441, 1410, 1340, 1279, 1231,1097, 1073, 1029, 953, 902, 773, 702 cm-1.

12f:

¹H NMR (500 MHz, CDCl3) δ 7.41-7.12 (m, 10H), 5.07 (d, J=1.4 Hz, 1H),4.99 (d, J=1.5 Hz, 1H), 3.99-3.92 (s, 1H), 2.38-2.26 (m, 5H), 2.11-1.98(m, 5H), 1.75-1.58 (m, 6H), 1.36-1.17 (m, 11H). ¹³C NMR (126 MHz, CDCl3)δ 179.38, 154.57, 144.15, 141.17, 139.10, 137.37, 129.69, 127.74,127.71, 127.62, 126.65, 126.59, 114.99, 82.11, 69.33, 55.69, 40.23,33.99, 33.94, 32.08, 29.65, 29.57, 29.40, 29.33, 29.19, 29.03, 27.76,24.68. LRMS (ESI, APCI) m/z: calc'd for C33H41O3 [M−H]− 485.3, found485.3. Calc'd for C33H41O2 [M−OH]+469.3, found 468.9 HRMS (ESI) m/z:calc'd for C33H41O3 [M−H]− 485.30612, found 485.30646 FT-IR (neat):1352, 3079, 3052, 3019, 2925, 2853, 1708, 1598, 1491, 1441, 1410, 1340,1281, 1230, 1096, 1073, 1029, 954, 902, 773, 702 cm-1.

Synthetic Scheme for [3.3.0] Bicyclic Phosphorylcholines

General Procedure for Phosphorylcholines

The required cyclized primary alcohols (5d-12d) (1.0 equiv.) weredissolved in toluene and cooled to 0° C. Triethylamine (2.0 equiv.) wasadded and the solution was briefly stirred. 2-chloro1,3,2,-dioxaphospholane 2-oxide (2.0 equiv.) was then added dropwise.The reaction mixture was stirred for 4 hours at which time completionwas detected by LCMS. The reaction mixture was filtered through a cottonplug to remove the majority of the resulting triethylammonium salts,concentrated in vacuo, and reacted without further purification. Therequired cyclized phospholane intermediates (1.0 equiv.) were dissolvedin acetonitrile, placed in a pressure tube open to air, equipped a stirbar and frozen at −78° C. Trimethylamine (excess) was condensed into thecold pressure tube, which was then capped and allowed to warm to roomtemperature before heating to 90° C. for 16 h. After reacting for 16 h,the reaction mixture was cooled to −78° C., uncapped, and allowed towarm to room temperature. The mixture was concentrated in vacuo andcarried on without further purification.

Crude material from the previous reaction was dissolved in acetonitrile.Concentrated HCl (5-10 equiv.) was added and the mixture was allowed tostir for 30 minutes at room temperature. When the reaction was complete,as determined by LCMS, the reaction mixture was concentrated in vacuoand subjected to preparatory HPLC to isolate the desired product as themajor exo diastereomer.

Identi- fier Compound Name  5g4-(6-exo-hydroxy-3-phenyl-3a-(1-phenylvinyl)-1,3a,4,5,6,6a-hexahydropentalen-2-yl)butyl (2-(trimethylammonio)ethyl) phosphate  6g5-(6-exo-hydroxy-3-phenyl-3a-(1-phenylvinyl)-1,3a,4,5,6,6a-hexahydropentalen-2-yl)pentyl (2-(trimethylammonio)ethyl) phosphate  7g6-(6-exo-hydroxy-3-phenyl-3a-(1-phenylvinyl)-1,3a,4,5,6,6a-hexahydropentalen-2-yl)hexyl(2-(trimethylammonio)ethyl) phosphate  8g7-(6-exo-hydroxy-3-phenyl-3a-(1-phenylvinyl)-1,3a,4,5,6,6a-hexahydropentalen-2-yl)heptyl (2-(trimethylammonio)ethyl) phosphate  9g8-(6-exo-hydroxy-3-phenyl-3a-(1-phenylvinyl)-1,3a,4,5,6,6a-hexahydropentalen-2-yl)octyl (2-(trimethylammonio)ethyl) phosphate 10g9-(6-exo-hydroxy-3-phenyl-3a-(1-phenylvinyl)-1,3a,4,5,6,6a-hexahydropentalen-2-yl)nonyl (2-(trimethylammonio)ethyl) phosphate 11g10-(6-exo-hydroxy-3-phenyl-3 a-(1-phenylvinyl)-1,3a,4,5,6,6a-hexahydropentalen-2-yl)decyl (2-(trimethylammonio)ethyl) phosphate 12g11-(6-exo-hydroxy-3-phenyl-3a-(1-phenylvinyl)-1,3a,4,5,6,6a-hexahydropentalen-2-yl)undecyl (2-(trimethylammonio)ethyl) phosphateCharacterization Data for Phosphorylcholines (5g-12g)5g:

¹H NMR (500 MHz, CDCl₃) δ 7.28-7.13 (m, 10H), 5.02 (s, 1H), 4.99 (s,1H), 4.19 (s, 2H), 3.81 (s, 1H), 3.76 (s, 2H), 3.64 (s, 2H), 3.21 (s,9H), 2.26-2.05 (m, 3H), 2.01-1.87 (m, 1H), 1.68-1.36 (m, 9H). 31P NMR(121 MHz, CDCl₃) δ −0.76. LRMS (ESI, APCI) m/z: calc'd for C31H43O5NP[M+H]+: 540.3, found 540.3

6g:

¹H NMR (600 MHz, CDCl₃) δ 7.31-7.24 (m, 4H), 7.25-7.14 (m, 6H), 4.99 (s,2H), 4.19 (s, 2H), 3.86 (s, 1H), 3.75 (s, 2H), 3.63 (s, 2H), 3.20 (s,9H), 2.25-2.18 (m, 1H), 2.16-2.04 (m, 2H), 1.95-1.87 (m, 1H), 1.70-1.60(m, 1H), 1.60-1.52 (m, 4H), 1.53-1.43 (m, 1H), 1.42-1.30 (m, 3H),1.31-1.18 (m, 2H). ³¹P NMR (121 MHz, CDCl3) δ −0.82. LRMS (ESI, APCI)m/z: calc'd for C32H45O5NP [M+H]+: 554.3, found 554.2

7g:

¹H NMR (500 MHz, CDCl3) δ 7.41-7.26 (m, 4H), 7.28-7.12 (m, 6H), 5.03 (s,1H), 5.01 (s, 1H), 4.24 (s, 2H), 3.88 (s, 1H), 3.80-3.67 (m, 4H), 3.28(s, 9H), 2.28-2.05 (m, 3H), 2.01-1.80 (m, 1H), 1.72-1.49 (m, 3H),1.40-0.82 (m, 10H). ³¹P NMR (121 MHz, Chloroform-d) 6-0.59. LRMS (ESI,APCI) m/z: calc'd for C33H47O5NP [M+H]+ 568.3, found 568.2

8g:

¹H NMR (500 MHz, CDCl3) δ 7.37-7.11 (m, 10H), 5.02 (s, 1H), 4.99 (s,1H), 4.21 (s, 2H), 3.87 (s, 1H), 3.77 (s, 2H), 3.67 (s, 1H), 3.25 (s,9H), 2.29-2.16 (m, 1H), 2.15-2.02 (m, 2H), 2.00-1.90 (m, 1H), 1.72-1.57(m, 3H), 1.57-1.48 (m, 3H), 1.37-1.14 (m, 9H). 13C NMR (300 MHz, CDCl3)δ 154.59, 144.07, 140.89, 139.62, 137.37, 129.56, 127.83, 127.67,126.64, 114.74, 81.46, 69.08, 66.14, 65.72, 59.20, 55.70, 54.25, 39.99,34.18, 32.02, 30.77, 29.47, 29.31, 29.17, 27.57, 25.56. 31P NMR (300MHz, CDCl3) δ −0.51. HRMS (ESI) m/z: calc'd for C34H49O5NP [M+H]+:582.33429, found 582.33380 IR (cm-1): 3373 (b), 2930, 2854, 1709, 1668,1598, 1490, 1440, 1343, 1227, 1090.

9g:

¹H NMR (600 MHz, CDCl3) δ 7.33-7.15 (m, 10H), 5.00 (s, 1H), 4.96 (s,1H), 4.22 (s, 2H), 3.89 (s, 1H), 3.81-3.75 (m, 2H), 3.74-3.66 (m, 2H),3.27 (s, 9H), 2.28-2.17 (m, 2H), 2.10-2.04 (m, 1H), 1.96-1.87 (m, 1H),1.71-1.58 (m, 3H), 1.57-1.48 (m, 2H), 1.36-1.13 (m, 12H). ¹³C NMR (151MHz, CDCl3) δ 154.59, 144.14, 141.04, 139.50, 137.40, 129.61, 127.78,127.63, 126.61, 126.55, 114.80, 81.66, 69.23, 66.29, 59.11, 55.61,54.37, 53.41, 40.05, 37.14, 34.27, 32.02, 30.82, 29.35, 29.17, 28.91,28.84, 27.47, 25.57, 22.61. ³¹P NMR (300 MHz, CDCl3) δ −0.43. LRMS (ESI,APCI) m/z: calc'd for C35H51O5NP [M+H]+ 596.3, found 596.3. HRMS (ESI)m/z: calc'd for C35H51O5NP [M+H]+: 596.34994, found 596.3939 IR (cm-1):3355 (b), 2927, 2854, 2187, 1669, 1491, 1440, 1227, 1090.

10g:

¹H NMR (600 MHz, CDCl3) δ 7.32-7.14 (m, 10H), 5.01 (d, J=7.6 Hz, 1H),4.95 (d, J=7.9 Hz, 1H), 4.20 (s, 2H), 3.87 (s, 1H), 3.75 (s, 3H), 3.67(s, 3H), 3.25 (s, 9H), 2.27-2.18 (m, 2H), 2.06-1.98 (m, 2H), 1.94 (p,J=7.0 Hz, 1H), 1.70-1.58 (m, 4H), 1.58-1.48 (m, 3H), 1.36-1.10 (m, 12H).¹³C NMR (126 MHz, CDCl3) δ 154.65, 144.17, 141.06, 139.41, 137.41,129.65, 127.76, 127.67, 127.62, 126.63, 126.57, 114.85, 81.72, 69.29,66.24, 65.92, 59.30, 55.66, 54.36, 40.17, 34.15, 32.09, 30.89, 29.51,29.28, 29.23, 27.66, 27.61, 25.76. 31P NMR (121 MHz, CDCl3) δ −0.73.LRMS (ESI, APCI) m/z: calc'd for C36H53O5NP [M+H]+ 610.4, found 609.8.HRMS (ESI) m/z: calc'd for C36H53O5NP [M+H]+: 610.36559, found 610.36552IR (cm-1): 3372 (b), 2926, 2853, 1653, 1491, 1440, 1228, 1090

11g:

¹H NMR (600 MHz, CDCl3) δ 7.34-7.14 (m, 10H), 5.03 (d, J=4.9 Hz, 1H),4.95 (d, J=5.0 Hz, 1H), 4.24 (s, 2H), 3.90 (s, 1H), 3.79 (q, J=6.4 Hz,2H), 3.74 (s, 2H), 3.28 (s, 9H), 2.34-2.23 (m, 2H), 2.10-1.92 (m, 3H),1.64 (q, J=10.2, 6.5 Hz, 2H), 1.56 (t, J=7.3 Hz, 2H), 1.37-1.10 (m,16H). ¹³C NMR (126 MHz, CDCl3) δ 154.71, 144.23, 141.07, 139.40, 137.43,129.69, 127.73, 127.70, 127.60, 126.63, 126.56, 81.67, 69.33, 66.20,65.87, 59.29, 55.64, 54.31, 40.23, 34.08, 32.18, 30.91, 30.86, 29.55,29.32, 29.22, 29.17, 27.60, 25.82. ³¹P NMR (121 MHz, CDCl3) δ −0.75.LRMS (ESI, APCI) m/z: calc'd for C37H55O5NP [M+H]+: 624.4, found 624.3

12g:

¹H NMR (600 MHz, CDCl3) δ 7.35-7.15 (m, 12H), 5.03 (s, 1H), 4.96 (s,1H), 4.26 (s, 2H), 3.91 (s, 1H), 3.85-3.77 (m, 2H), 3.75 (s, 2H), 3.29(s, 9H), 2.32 (dd, J=16.7, 9.4 Hz, 1H), 2.26 (d, J=9.6 Hz, 1H),2.10-1.94 (m, 4H), 1.71-1.61 (m, 3H), 1.61-1.53 (m, 2H), 1.38-1.14 (m,16H). ³¹P NMR (121 MHz, CDCl3) δ −0.90. ¹³C NMR (126 MHz, CDCl3) δ154.72, 144.22, 141.10, 139.32, 137.43, 129.69, 114.91, 81.70, 69.33,66.20, 65.88, 59.31, 55.67, 54.37, 40.25, 34.02, 32.18, 31.58, 30.91,30.86, 29.60, 29.51, 29.43, 29.40, 29.36, 29.34, 29.24, 27.65, 25.76,22.65. LRMS (ESI, APCI) m/z: calc'd for C38H57O5NP [M+H]+ 638.4, found637.8 HRMS (ESI) m/z: calc'd for C38H57O5NP [M+H]+: 638.39689, found638.39741 FT-IR (neat): 3362 (b), 2924, 2853, 1667, 1490, 1440, 1227,1090 cm-1.

Synthetic Scheme for Locked Synthetic Phospholipid Mimic (13b)

1-((R)-3-(hydroxymethyl)pyrrolidin-1-yl)-8-(6-(exo)(methoxymethoxy)-3-phenyl-3a-(1-phenylvinyl)-1,3a,4,5,6,6a-hexahydropentalen-2-yl)octan-1-one(13a)

In a reaction tube equipped with a stir bar, 8d was dissolved inacetonitrile. Tetrapropylammonium perruthenate (TPAP, 0.1 equiv.),N-Methylmorpholine-N-Oxide (NMO, 10.0 equiv.), and water (10.0 equiv.)were added. The reaction mixture was allowed to stir until complete byTLC and LCMS, 1-16 h. Upon reaction completion, the reaction mixture wasconcentrated and subjected directly to silica for purification in 20-50%EtOAc/Hex (0.1% Acetic acid) to afford the desired compound as a clear,colorless oil (96%)

This resulting carboxylic acid was dissolved in DMF in a reaction tubeequipped with a stir bar.(2-(1H-benzotriazol-1-yl)-1,1,3,3-tetramethyluronium hexafluorophosphate(HBTU, 1.2 equiv.) was added. (3R)-pyrrolidin-3-ylmethanol (1.2 equiv.)was added in DMF before diisopropylethylamine (2.0 equiv.) was added.The reaction mixture was heated and allowed to stir at 60° C. for 1 h,until complete by LCMS. The mixture was cooled to room temperaturebefore being poured into water and extracted with EtOAc. The combinedorganic layers were dried with MgSO₄ and concentrated before beingpurified by silica gel chromatography in 100% EtOAc/Hex to afford thedesired compound 13a as a clear, colorless oil (99%).

((3R)-1-(8-(6-(exo)-hydroxy-3-phenyl-3a-(1-phenylvinyl)-1,3a,4,5,6,6a-hexahydropentalen-2-yl)octanoyl)pyrrolidin-3-yl)methyl(2-(trimethylammonio)ethyl) phosphate (13b)

In a scintillation vial equipped with a stir bar, 13a was dissolved intoluene. 2-Chloro-1,3,2-dioxaphospholane 2-oxide (2.0 equiv.) was added,followed by triethylamine (2.0 equiv.) The resulting reaction mixturewas stirred for 1-4 h, until reaction was complete by TLC. The resultingmixture containing the cyclic phospholane intermediate was filtered overa cotton plug to remove excess ammonium salts and concentrated. Thecrude reaction mixture was taken up in acetonitrile and transferred intoa pressure tube and cooled to −78° C. Trimethylamine (neat, 2-5 mL) wascondensed into the pressure tube at −78° C. The tube was capped, allowedto warm to room temperature, and then heated to 90° C. for 16 h. Afterthe reaction was complete, the pressure tube was allowed to cool to R.T.before being further cooled to −78° C. and uncapped. The solution wasallowed to re-warm to room temperature before being concentrated insidea fume hood and carried on without further purification. The crudereaction mixture was dissolved in acetonitrile, and 2-5 drops ofconcentrated HCl was added. The mixture was allowed to stir until thereaction was complete (5-30 min, monitored by LCMS), and thenconcentrated. The crude reaction mixture was purified on alumina in65/35/5 CH2Cl2/MeOH/NH4OH to afford the desired compound 13b.

¹H NMR (600 MHz, CD3OD) δ 7.37-7.14 (m, 10H), 4.97 (d, J=1.6 Hz, 1H),4.94 (d, J=1.7 Hz, 1H), 4.27 (s, 2H), 3.94-3.88 (m, 1H), 3.87 (s, 1H),3.85-3.80 (m, 1H), 3.69-3.64 (m, 2H), 3.63-3.47 (m, 2H), 3.34 (s, 2H),3.24 (s, 9H), 2.63 (p, J=7.1 Hz, 1H), 2.54 (p, J=7.1 Hz, 1H), 2.35-2.24(m, 5H), 2.14-2.02 (m, 2H), 2.01 (t, J=8.7, 6.7 Hz, 2H), 1.90-1.80 (m,1H), 1.79-1.64 (m, 1H), 1.68-1.59 (m, 2H), 1.59-1.45 (m, 2H), 1.40-1.17(m, 7H). ¹³C NMR (151 MHz, CD3OD) δ 172.47, 155.17, 144.09, 140.82,139.49, 137.35, 129.41, 127.54, 127.26, 126.34, 113.56, 81.37, 69.28,66.46, 66.18, 66.02, 59.02, 55.08, 53.28, 45.99, 44.89, 39.84, 34.02,33.82, 33.12, 31.72, 29.13, 29.10, 28.95, 28.91, 28.79, 28.75, 27.80,27.30, 27.28, 26.19, 24.60, 24.59. LRMS (ESI, APCI) m/z: calc'd forC40H58N2O6P [M+H]+: 693.4, found 693.6 HRMS (ESI) m/z: calc'd forC40H58N2O6P [M+H]+ 693.40300, found 693.40270.

Synthetic Procedure for Phospholipid Mimic 14d

The synthetic procedure for preparing compound 14d is provided for inFIG. 7.

(R)-3-((tert-butyldiphenylsilyl)oxy)-2-hydroxypropyl8-(6-(exo)-(methoxymethoxy)-3-phenyl-3a-(1-phenylvinyl)-1,3a,4,5,6,6a-hexahydropentalen-2-yl)octanoate(14a)

In a scintillation vial equipped with stir bar, 8d was dissolved inacetonitrile. Tetrapropyl ammonium perruthenate (TPAP, 0.1 equiv.),N-Methylmorpholine-N-Oxide (NMO, 10.0 equiv.), and water (10.0 equiv.)were added. The reaction mixture was allowed to stir until complete byTLC and LCMS, 1-16 h. Upon reaction completion, the reaction mixture wasconcentrated and subjected directly to silica for purification in 20-50%EtOAc/Hex (0.1% Acetic acid) to afford the desired compound as a clear,colorless oil (96%). In a reaction tube equipped with a stir bar,(R)-tert-butyl(oxiran-2-ylmethoxy)diphenylsilane was dissolved indiethyl ether. Co[Salen](S,S)-(+)-N,N′bix(3,5-di-tert-butylsalicylidene)-1,2-cyclohexanediamonocobalt (II) (0.01 equiv.) was added and the reaction was allowed to stirfor 1 h open to air to activate. After 1 h, the diethylether wasevaporated. The previously prepared carboxylic acid (1.0 equiv.) anddiisopropyl ethyl amine (1.0 equiv.) was added to the reaction tube andthe reaction was allowed to stir, neat, at room temperature for 3-16 h,until complete by TLC and LCMS. When reaction was determined to becomplete, the crude reaction mixture was directly purified by silica gelchromatography in 5-10% EtOAc/Hex to afford the desired compound 14a(28%).

(R)-3-((tert-butyldiphenylsilyl)oxy)-2-(propionyloxy)propyl8-(6-(exo)-(methoxymethoxy)-3-phenyl-3a-(1-phenylvinyl)-1,3a,4,5,6,6a-hexahydropentalen-2-yl)octanoate(14b)

In a scintillation vial equipped with a stir bar, 14a was dissolved inDCM. Propionyl chloride (4.0 equiv.) was added at room temperature,followed by triethylamine (4.0 equiv.). The reaction was allowed to stiruntil complete by TLC (4 h). The resulting reaction mixture wasconcentrated and purified by silica gel chromatography in 5-10%EtOAc/Hex to afford the desired compound 14b (74%).

(S)-3-hydroxy-2-(propionyloxy)propyl8-(6-(exo)-(methoxymethoxy)-3-phenyl-3a-(1-phenylvinyl)-1,3a,4,5,6,6a-hexahydropentalen-2-yl)octanoate(14c)

In a scintillation vial equipped with a stir bar, 14b was dissolved inTHF. Tetrabutylammoniumfluoride (1.1 equiv.) was added, and the reactionmixture was allowed to stir for 16 h. The resulting mixture wasconcentrated and purified by silica gel chromatography in 20-50%EtOAc/Hex to afford the desired compound 14c (>99%).

(2R)-3-((8-(6-(exo)-hydroxy-3-phenyl-3a-(1-phenylvinyl)-1,3a,4,5,6,6a-hexahydropentalen-2-yl)octanoyl)oxy)-2-(propionyloxy)propyl(2-(trimethylammonio)ethyl) phosphate (14d)

In a scintillation vial equipped with a stir bar, 14c was dissolved intoluene. 2-Chloro-1,3,2-dioxaphospholane 2-oxide (2.0 equiv.) was added,followed by triethylamine (2.0 equiv.) The resulting reaction mixturewas stirred for 1-4 h, until reaction was complete by TLC. The resultingmixture containing the cyclic phospholane intermediate was filtered overa cotton plug to remove excess ammonium salts and concentrated. Thecrude reaction mixture was taken up in acetonitrile and transferred intoa pressure tube and cooled to −78° C. Trimethylamine (neat, 2-5 mL) wascondensed into the pressure tube at −78° C. The tube was capped, allowedto warm to room temperature, and then heated to 90° C. for 16 h. Afterthe reaction was complete, the pressure tube was allowed to cool to R.T.before being further cooled to −78° C. and uncapped. The solution wasallowed to re-warm to room temperature before being concentrated insidea fume hood and carried on without further purification. The crudereaction mixture was dissolved in acetonitrile, and 2-5 drops ofconcentrated HCl was added. The mixture was allowed to stir until thereaction was complete (5-30 min, monitored by LCMS), and thenconcentrated. The crude reaction mixture was purified via preparatoryHPLC to afford the desired compound 14d.

¹H NMR (600 MHz, CDCl₃) δ 7.37-7.15 (m, 10H), 5.04 (s, 1H), 4.98 (s,1H), 4.51 (s, OH), 4.44-4.28 (m, 4H), 4.28-4.17 (m, 2H), 4.18-4.06 (m,1H), 3.92 (s, 1H), 3.80-3.69 (m, 2H), 3.30 (s, 9H), 2.38-2.22 (m, 6H),2.12-1.93 (m, 4H), 1.60-1.48 (m, 4H), 1.38-1.13 (m, 9H), 1.10 (t, J=7.5Hz, 3H). LRMS (ESI, APCI) m/z: calc'd for C41 [M]+739.4, found 739.6HRMS (ESI) m/z: calc'd for C41H59O9NP [M+H]+ 740.39393, found 740.39220.Calc'd for C41H48O9NPNa [M+Na]+762.37569, found 762.37414. FT-IR (neat):3357, 3079, 3020, 2925, 2854, 1736, 1715, 1686, 1618, 1598, 1555, 1490,1463, 1440, 1363, 1250, 1201, 1175, 1085, 1025, 967, 912, 800, 760, 702cm-1.

Synthetic Scheme for Internal Styrene Modifications (15f-18f, 15g-18g).

The synthetic procedure for preparing compounds 15f-18f, 15g-18g isprovided for in FIG. 1D.

General Sonogashira Coupling Procedure (15a-18b)

To an oven-dried round bottom flask was addedbis(triphenylphosphine)palladium dichloride (0.03 equiv.) and copperiodide (0.06 equiv.). Triethylamine was added to make a 1.0 M solutionbefore the addition of required aryl halide (1.0 equiv.) The resultingyellow mixture was sparged by bubbling the solution with nitrogen for aperiod of 30 minutes, at which point 4-pentyl-1-ol (1.2 equiv.) wasadded portionwise and the sparging needle was replaced with a nitrogeninlet. The solution rapidly darkened and formed a slurry, and was heatedat 60° C. for 2 hours, at which point the reaction was complete by TLC.The resulting black solution was cooled and ether was added toprecipitate a black solid. The resulting slurry was filtered over a plugof celite and eluted with ether. The filtrate was concentrated in vacuoto afford a reddish-brown oil, which was then purified on silica in 30%EtOAc/hex to afford a yellow oil.

5-(3-bromophenyl)pent-4-yn-1-ol (15a)

The general Sonogashira coupling procedure was followed using1-bromo-3-iodobenzene as the aryl halide. The compound was purified in10-30% EtOAc/Hex (3.1 g 92%).

5-(5-bromo-2-fluorophenyl)pent-4-yn-1-ol (16a)

The general Sonogashira coupling procedure was followed using4-bromo-1-fluoro-2-iodobenzne as the aryl halide. The compound waspurified in 10-30% EtOAc/Hex (3.9 g, 75%).

5-(5-bromo-2-methylphenyl)pent-4-yn-1-ol (17a)

The general Sonogashira coupling procedure was followed using4-methyl-2-iodo-1-methylbenzene as the aryl halide. The compound waspurified in 10-30% EtOAc/Hex as the eluent, (3.6 g, 81%).

5-(3-bromo-5-fluorophenyl)pent-4-yn-1-ol (18a)

The general Sonogashira coupling procedure was followed using1-bromo-3-fluoro-5-iodobenzene as the aryl halide. The compound waspurified in 20% EtOAc/Hex (3.5 g 90%).

General Swern Oxidation Procedure (15b-18b)

To an oven-dried three-neck roundbottom flask charged with a stir barwas added oxalyl chloride (1.1 equiv.) in DCM (0.1M). The solution wascooled to −78° C. before the dropwise addition of dimethylsulfoxide (1.3equiv.) in DCM. After effervescence ceased, the required alcohol (1.0equiv.) was added dropwise in DCM. The reaction mixture was stirred at−78° C. for 1.5 h before quenching with triethylamine (2.5 equiv.) andallowing to warm to room temperature before additional quenching withsaturated ammonium chloride. The reaction mixture was then poured overwater and extracted with DCM, dried with MgSO₄, concentrated, andpurified by silica gel chromatography to afford a (typically paleyellow) oil.

5-(3-bromophenyl)pent-4-ynal (15b)

The general procedure for Swern oxidation was followed, using5-(3-bromophenyl)pent-4-yn-1-ol as the alcohol. The crude oil waspurified on silica gel with 10-50% EtOAc/Hex (3.4 g, 81%).

5-(5-bromo-2-fluorophenyl)pent-4-ynal (16b)

The general procedure for Swern oxidation was followed, using5-(5-bromo-2-fluorophenyl)pent-4-yn-1-ol as the alcohol. The crude oilwas purified on silica gel with 10-20% EtOAc/Hex (2.4 g, 62%).

5-(5-bromo-2-methylphenyl)pent-4-ynal (17b)

The general procedure for Swern oxidation was followed using5-(5-bromo-2-methylphenyl)pent-4-yn-1-ol as the alcohol. The crude oilwas purified on silica gel with 10% EtOAc/Hex (0.6 g, 17%).

5-(3-bromo-5-fluorophenyl)pent-4-ynal (18b)

The general procedure for Swern oxidation was followed, using5-(3-bromo-5-fluorophenyl)pent-4-yn-1-ol as the alcohol. The crude oilwas purified on silica gel with 10-20% EtOAc/Hex, (3.5 g, 90

General Procedure for Grignard Addition (15c-18c)

To an oven-dried 3-neck flask equipped with a stir bar was added therequired aldehyde (1.0 equiv.) in THF. The solution was cooled to −78°C. before the addition of vinylmagnesium bromide (1.5 equiv.). Thereaction was stirred and allowed to warm to room temperature overnightbefore quenching with saturated ammonium chloride. The reaction mixturewas poured over water and extracted with ethyl acetate, dried withMgSO₄, and concentrated before being purified by silica gelchromatography.

7-(3-bromophenyl)hept-1-en-6-yn-3-ol (15c)

The general procedure for Grignard addition was followed, using5-(3-bromophenyl)pent-4-ynal as the aldehyde. The crude oil was thenpurified on silica gel with 5-10% EtOAc/Hex, (1.12 g, 55%).

7-(5-bromo-2-fluorophenyl)hept-1-en-6-yn-3-ol (16c)

The general procedure for Grignard addition was followed, using5-(5-bromo-2-fluorophenyl)pent-4-ynal as the aldehyde. The crude oil wasthen purified on silica gel with 20% EtOAc/Hex, (3.3 g, 90%)

7-(5-bromo-2-methylphenyl)hept-1-en-6-yn-3-ol (17c)

The general procedure for Grignard addition was followed, using5-(5-bromo-2-methylphenyl)pent-4-ynal as the aldehyde. The crude oil wasthen purified on silica gel with 20% EtOAc/Hex, (1.4 g, 60%).

7-(3-bromo-5-fluorophenyl)hept-1-en-6-yn-3-ol (18c)

The general procedure for Grignard addition was followed, using5-(3-bromo-5-fluorophenyl)pent-4-yn-1-ol as the aldehyde. The crude oilwas then purified on silica gel with 20% EtOAc/Hex, (3.0 g, 81%).

General Protection Procedures and Characterization Data for ProtectedEnynes (15d-18d, 19)

Tert-butyldimethyl((7-phenylhept-1-en-6-yn-3-yl)oxy)silane (19): In anoven-dried three-neck flask equipped with stir bar and flushed withnitrogen was added imidazole (4.0 equiv.) and 4-dimethylaminopyridine(2.0 equiv.). After further evacuation and backfilling with nitrogen,the solids were dissolved in dry THE and cooled to −78° C. Followingaddition of a solution of the alcohol (1.0 equiv.) in THF,tert-butyldimethylsilyl triflate (2.0 equiv.) was added dropwise, andallowed to stir for 6 hours. The reaction was then quenched by saturatedaqueous NH₄Cl, and extracted with ether (3×); the combined organiclayers were washed with brine, dried over MgSO₄, filtered andconcentrated in vacuo. The residue was purified by silica gelchromatography in 5-10% EtOAc/Hex to afford a clear yellow oil (8.0 g,99% yield). The spectral data reported are consistent with literature.

General Procedure for Alcohol Protection with Methoxymethyl (MOM) Ether(15d-18d)

The required unprotected enyne (15c-18c) (1.0 equiv.) was dissolved inDCM, followed by diisopropylethyl amine (1.25 equiv.) Chloromethylmethyl ether (1.5 equiv.) was added and the reaction mixture was stirredat 30° C. until completion (typically 1-4 hours). The reaction mixturewas cooled to room temperature before being poured onto water, washedwith dilute HCl (1 M), and extracted with DCM. The organic layers weredried with MgSO₄, filtered, and concentrated before being subjected tosilica gel chromatography.

1-bromo-3-(5-(methoxymethoxy)hept-6-en-1-yn-1-yl)benzene (15d)

The general procedure for MOM ether protection was followed, using7-(3-bromophenyl)hept-1-en-6-yn-3-ol as the enyne. The crude oil waspurified in 5% EtOAc/Hex (2.1 g, 92%).

4-bromo-1-fluoro-2-(5-(methoxymethoxy)hept-6-en-1-yn-1-yl)benzene (16d)

The general procedure for MOM ether protection was followed, using7-(5-bromo-2-fluorophenyl)hept-1-en-6-yn-3-ol as the enyne. The crudeoil was purified in 2-10% EtOAc/Hex (1.1 g, 88%).

4-bromo-2-(5-(methoxymethoxy)hept-6-en-1-yn-1-yl)-1-methylbenzene (17d)

The general procedure for MOM ether protection was followed, using7-(5-bromo-2-methylphenyl)hept-1-en-6-yn-3-ol as the enyne. The crudeoil was purified in 2-10% EtOAc/Hex (0.2 g, 78%).

1-bromo-3-fluoro-5-(5-(methoxymethoxy)hept-6-en-1-yn-1-yl)benzene (18d)

The general procedure for MOM ether protection was followed, using7-(3-bromo-5-fluorophenyl)hept-1-en-6-yn-3-ol as the enyne. The crudeoil was purified in 2-10% EtOAc/Hex (1.0 g, 77%).

General Procedure for Zirconocene Mediated Cyclization and MOMDeprotection (15e-18e)

Bis(cyclopentadienyl)zirconium(IV) dichloride (zirconecene dichloride)(1.2 equiv.) was dried by azeotroping away latent water with benzenefour times before being placed under nitrogen, dissolved intetrahydrofuran (THF) and cooled to −78° C. in a dry ice/acetone bath.The resulting solution of zirconecene dichloride was treated with nBuLi(2.4 equiv.) to form a clear, light yellow solution and allowed to stir.After approximately 30 minutes, azeotroped required enyne (1.0 equiv.)in dry THF was added portionwise to afford a red-orange solution, andthe reaction mixture was held at −78° C. for 30 minutes before allowingto warm to room temperature and stir over 2.5 hours. The reactionmixture was then re-cooled to −78° C. and 1,1-dibromoheptane (1.1equiv.) were added in dry THF. Freshly prepared lithium diisopropylamine(LDA, 1.0 M, 1.1 equiv.) was added at −78° C. and stirred for 15minutes. Lithium phenylacetylide (3.6 equiv.) was then prepared andadded to the reaction mixture dropwise in dry THF. The resulting darkreddish brown solution was stirred at −78° C. for one hour. The reactionwas then quenched with methanol and saturated aqueous sodium bicarbonateand allowed to warm to room temperature to form a light yellow slurry.The resulting slurry was poured over water and extracted with ethylacetate four times. The combined organic layers were washed with brine,dried with MgSO₄, and concentrated in vacuo. The resulting colored oil(typically yellow, orange, or brown), was roughly purified on a plug ofsilica and eluted with 20% EtOAc/Hex to afford a yellow oil which is amixture of phenylacetylene, desired protected [3.3.0] bicycliccompounds, and (in some cases) protected protonolysis byproduct. Thisoil was carried on without further purification. This procedure affordsexo and endo diastereomers in a 7:1 ratio as determined bycharacteristic 1H NMR signals.

The crude mixture (1.0 equiv.) was dissolved in acetonitrile and a fewdrops of concentrated HCl (excess) was added. The resulting darkblue-purple solution was stirred open to air for approximately 30minutes (monitored via LCMS), concentrated, and subjected directly tosilica gel chromatography in 5-20% EtOAc/Hexanes to afford the desiredcompound as a pale yellow oil.

(exo)-4-(3-bromophenyl)-5-hexyl-3a-(1-phenylvinyl)-1,2,3,3a,6,6a-hexahydropentalen-1-ol(15e)

The general procedure for zirconecene mediated cyclization and MOMdeprotection was followed, using1-bromo-3-(5-(methoxymethoxy)hept-6-en-1-yn-1-yl)benzene as the enyne.The crude oil was purified by silica gel chromatography in 5-20%EtOAc/Hex(exo)-4-(5-amino-2-fluorophenyl)-5-hexyl-3a-(1-phenylvinyl)-1,2,3,3a,6,6a-hexahydropentalen-1-ol(16e)

The general procedure for zirconecene mediated cyclization and MOMdeprotection was followed, using4-bromo-1-fluoro-2-(5-(methoxymethoxy)hept-6-en-1-yn-1-yl)benzene as theenyne. The crude oil was purified by silica gel chromatography in 5-20%EtOAc/Hex, affording both the desired compound and lithium-halogenexchange byproduct in an appreciable amount.

This mixture was carried on without further purification.

(exo)-4-(5-bromo-2-methylphenyl)-5-hexyl-3a-(1-phenylvinyl)-1,2,3,3a,6,6a-hexahydropentalen-1-ol(17e)

The general procedure for zirconecene mediated cyclization and MOMdeprotection was followed, using4-bromo-2-(5-(methoxymethoxy)hept-6-en-1-yn-1-yl)-1-methylbenzene as theenyne. The crude oil was purified by silica gel chromatography in 5-20%EtOAc/Hex.

(exo)-4-(3-bromo-5-fluorophenyl)-5-hexyl-3a-(1-phenylvinyl)-1,2,3,3a,6,6a-hexahydropentalen-1-ol(18e)

The general procedure for zirconecene mediated cyclization and MOMdeprotection was followed, using1-bromo-3-fluoro-5-(5-(methoxymethoxy)hept-6-en-1-yn-1-yl)benzene as theenyne. The crude oil was purified by silica gel chromatography in 5-10%EtOAc/Hex, affording both the desired compound and lithium-halogenexchange byproduct in an appreciable amount. This mixture was carried onwithout further purification.

General Procedure for Hydroxyl Coupling (15f-18f)

Potassium Hydroxide (3.0 equiv.),tris(dibenzylideneacetone)dipalladium(0) (0.01 equiv.), and tBuXPhos(0.04 equiv.) were placed in a reaction tube, which was evacuated andbackfilled with nitrogen three times. The solids were then suspended indegassed 1,4-dioxane under nitrogen. The required brominated [3.3.0]bicycle was added in 1,4-dioxane. Water (˜10 equiv.) was added. Thereaction mixture was heated to 80° C. and stirred for 16 hours. Afterstirring, the mixture was poured over water and EtOAc, and the organicswere washed with water and brine to remove 1,4-dioxane. The combinedorganic layers were dried with MgSO₄, concentrated, and purified onsilica in 20% EtOAc/Hex.

(exo)-5-hexyl-4-(3-hydroxyphenyl)-3a-(1-phenylvinyl)-1,2,3,3a,6,6a-hexahydropentalen-1-ol(15f)

The general procedure for hydroxyl coupling was followed, using (exo)-6(3-bromophenyl)-5-hexyl-3-(methoxymethoxy)-6a-(1-phenylvinyl)-1,2,3,3a,4,6a-hexahydropentalen-1-olas the brominated [3.3.0] bicycle. The crude oil was purified by silicagel chromatography in 20% EtOAc/Hex.

¹H NMR (500 MHz, CDCl₃) δ 7.30 (d, J=41.7 Hz, 5H), 7.17 (t, J=7.8 Hz,1H), 6.75 (t, J=9.3 Hz, 2H), 6.69 (s, 1H), 5.07 (s, 1H), 5.02 (s, 1H),4.88 (s, 1H), 3.94 (s, 1H), 2.35 (dd, J=17.3, 8.0 Hz, 1H), 2.27 (d,J=9.4 Hz, 1H), 2.05 (dt, J=21.8, 6.9 Hz, 4H), 1.77-1.48 (m, 5H),1.35-1.16 (m, 8H), 0.86 (t, J=7.0 Hz, 3H). ¹³C NMR (126 MHz, CDCl₃) δ154.91, 154.57, 144.10, 141.44, 139.08, 138.57, 128.78, 127.72, 126.68,122.42, 116.44, 115.08, 113.58, 82.06, 69.30, 55.82, 40.24, 34.00,32.06, 31.66, 29.74, 29.39, 27.92, 27.78, 22.59, 14.08. LRMS (ESI, APCI)m/z: calc'd for C28H37O2 [M+H]+ 404.3, found 403.8 FTIR (neat): 3368,3080, 2955, 2928, 2858, 1690, 1598, 1580, 1448, 1200, 1070, 755, 690cm-1.

(exo)-4-(2-fluoro-5-hydroxyphenyl)-5-hexyl-3a-(1-phenylvinyl)-1,2,3,3a,6,6a-hexahydropentalen-1-ol(16f)

The general procedure for hydroxyl coupling was followed, using(exo)-4-(5-amino-2-fluorophenyl)-5-hexyl-3a-(1-phenylvinyl)-1,2,3,3a,6,6a-hexahydropentalen-1-olas the brominated [3.3.0] bicycle. The crude oil was purified by silicagel chromatography in 20% EtOAc/Hex.

¹H NMR (500 MHz, CDCl3 δ 7.39-7.35 (m, 2H), 7.31-7.27 (m, 3H), 6.93 (t,J=8.8 Hz, 1H), 6.70 (dt, J=8.6, 3.5 Hz, 1H), 6.65 (dd, J=5.6, 3.2 Hz,1H), 5.10 (d, J=1.2 Hz, 1H), 4.93 (d, J=1.3 Hz, 1H), 4.61 (s, 1H), 3.97(s, 1H), 2.52 (dd, J=17.3, 9.7 Hz, 1H), 2.30 (d, J=9.7 Hz, 1H),2.12-2.03 (m, 2H), 1.95 (t, J=7.8 Hz, 2H), 1.87-1.78 (m, 1H), 1.73 (dd,J=12.3, 6.3 Hz, 1H), 1.68 (dd, J=13.1, 5.9 Hz, 1H), 1.45-1.27 (m, 2H),1.29-1.16 (m, 6H), 0.86 (t, J=7.1 Hz, 3H). ¹³C NMR (126 MHz, CDCl3) δ156.08, 154.54, 154.20, 150.56, 144.43, 143.62, 127.89, 127.41, 126.77,117.66, 117.63, 115.85, 115.65, 115.51, 114.92, 114.85, 81.94, 69.60,55.39, 40.57, 33.57, 33.09, 31.63, 30.00, 29.39, 27.23, 22.59, 14.09.¹⁹F NMR (282 MHz, cdcl3) δ −124.47. LRMS (ESI, APCI) m/z: calc'd forC28H36FO2 [M+H]+ 422.3, found 421.9 FTIR (neat): 3358, 3081, 2955, 2929,2856, 1737, 1491, 1443, 1204, 772, 703 cm-1.

(exo)-5-hexyl-4-(5-hydroxy-2-methylphenyl)-3a-(1-phenylvinyl)-1,2,3,3a,6,6a-hexahydropentalen-1-ol(17f)

The general procedure for hydroxyl coupling was followed, using(exo)-4-(5-bromo-2-methylphenyl)-5-hexyl-3a-(1-phenylvinyl)-1,2,3,3a,6,6a-hexahydropentalen-1-olas the brominated [3.3.0] bicycle. The crude oil was purified by silicagel chromatography in 20% EtOAc/Hex.

¹H NMR (500 MHz, Chloroform-d) δ 7.46-7.39 (m, 2H), 7.31-7.27 (m, 3H),7.09 (d, J=8.3 Hz, 1H), 6.67 (dd, J=8.3, 2.8 Hz, 1H), 6.56 (d, J=2.8 Hz,1H), 5.20 (d, J=1.1 Hz, 1H), 4.88 (d, J=1.2 Hz, 1H), 4.52 (s, 1H), 3.99(s, 1H), 3.49 (s, 3H), 2.64 (dd, J=17.3, 10.2 Hz, 1H), 2.33 (d, J=9.5Hz, 1H), 2.10-1.99 (m, 3H), 1.94-1.77 (m, 3H), 1.71 (ddd, J=19.1, 13.3,6.6 Hz, 2H), 1.42-1.12 (m, 7H), 0.85 (t, J=7.1 Hz, 3H).

(exo)-4-(3-fluoro-5-hydroxyphenyl)-5-hexyl-3a-(1-phenylvinyl)-1,2,3,3a,6,6a-hexahydropentalen-1-ol(18f)

The general procedure for hydroxyl coupling was followed, using(exo)-4-(3-bromo-5-fluorophenyl)-5-hexyl-3a-(1-phenylvinyl)-1,2,3,3a,6,6a-hexahydropentalen-1-olas the brominated [3.3.0] bicycle. The crude oil was purified by silicagel chromatography in 20% EtOAc/Hex.

¹H NMR (500 MHz, CDCl3) δ 7.35-7.28 (m, 2H), 7.26 (d, J=6.2 Hz, 3H),6.55-6.45 (m, 3H), 5.09 (s, 1H), 5.04 (s, 1H), 4.94 (s, 1H), 3.93 (s,1H), 2.35 (dd, J=17.0, 9.3 Hz, 1H), 2.28 (d, J=9.5 Hz, 1H), 2.15-1.99(m, 4H), 1.73-1.64 (m, 3H), 1.38-1.18 (m, 6H), 0.90-0.83 (m, 3H). ¹³CNMR (126 MHz, CDCl3) δ 162.01, 156.16, 154.36, 143.86, 142.29, 137.75,127.79, 127.67, 126.79, 115.30, 112.38, 109.15, 108.98, 101.67, 101.48,81.99, 69.26, 55.79, 40.23, 34.00, 32.02, 31.64, 29.71, 29.39, 27.71,22.59, 14.08. ¹⁹F NMR (282 MHz, cdcl3) 6-112.75. LRMS (ESI, APCI) m/z:calc'd for C28H36FO2 [M+H]+ 423.3, found 422.9 FTIR (neat): 3340, 3075,2960, 2931, 1738, 1618, 1583, 1448, 1352, 1217, 1002, 668 cm-1.

General Procedure for Amination (15g-18g) A reaction tube (A) equippedwith a magnetic stir bar was charged with tBuBrettPhos (0.04 equiv.) andsodium tert butoxide (3.0 equiv.) before being evacuated and backfilledwith nitrogen. The required brominated [3.3.0] bicycle was added indioxane before ammonia in dioxane (10 equiv.) was added and allowed tostir for −15 minutes. During this period in a separate reaction tube(B), tBuBrettPhos precatalyst (0.04 equiv.) was added, and the tube wasevacuated and backfilled with nitrogen. The tBuBrettPhos precatalyst wasthen dissolved in dry, degassed dioxane. The solution of tBuBrettPhosprecatalyst in reaction tube B was transferred to the stirring reactionmixture in reaction tube A. The nitrogen inlet was removed and thereaction mixture was heated to 80° C. for 16 hours behind a blastshield. After stirring, the mixture was cooled to room temperature,diluted with water and extracted with EtOAc. The organics were washedwith water and brine to remove dioxane, dried over MgSO₄, andconcentrated in vacuo. The crude oil was purified by silica gelchromatography in (typically 20-30%) EtOAc/Hex.

(exo)-4-(3-aminophenyl)-5-hexyl-3a-(1-phenylvinyl)-1,2,3,3a,6,6a-hexahydropentalen-1-ol(15g)

The general procedure for amine coupling was followed, using (exo)-6-(3bromophenyl)-5-hexyl-3-(methoxymethoxy)-6a-(1-phenylvinyl)-1,2,3,3a,4,6a-hexahydropentalen-1-olas the brominated [3.3.0] bicycle. The crude oil was purified by silicagel chromatography in 20% EtOAc/Hex. ¹H NMR (500 MHz, CDCl3) δ 7.49-7.20(m, 5H), 7.09 (dd, J=9.6, 5.6 Hz, 1H), 6.61 (d, J=7.4 Hz, 2H), 6.55 (s,1H), 5.06 (d, J=3.9 Hz, 1H), 5.02 (d, J=4.2 Hz, 1H), 3.93 (s, 1H), 3.52(s, 2H), 2.32 (dt, J=13.6, 6.7 Hz, 1H), 2.25 (d, J=8.8 Hz, 1H), 2.04(ddd, J=21.8, 11.9, 4.7 Hz, 5H), 1.77-1.62 (m, 3H), 1.38-1.06 (m, 8H),0.86 (t, J=7.1 Hz, 3H). ¹³C NMR (126 MHz, CDCl3) δ 154.73, 145.58,144.21, 140.96, 139.04, 138.54, 128.45, 127.79, 127.68, 126.61, 120.51,116.40, 114.90, 113.57, 109.99, 82.13, 55.91, 40.20, 34.07, 32.02,31.69, 29.78, 29.41, 27.82, 22.60, 14.08. LRMS (ESI, APCI) m/z: calc'dfor C28H38NO [M+H]+ 403.3, found 402.9

(exo)-4-(5-amino-2-fluorophenyl)-5-hexyl-3a-(1-phenylvinyl)-1,2,3,3a,6,6a-hexahydropentalen-1-ol(16g)

The general procedure for amine coupling was followed, using(exo)-4-(5-amino-2-fluorophenyl)-5-hexyl-3a-(1-phenylvinyl)-1,2,3,3a,6,6a-hexahydropentalen-1-olas the brominated [3.3.0] bicycle. The crude oil was purified by silicagel chromatography in 30-50% EtOAc/Hex.

¹H NMR (500 MHz, CDCl₃) δ 7.38 (dd, J=6.7, 2.9 Hz, 2H), 7.32-7.25 (m,2H), 6.99-6.79 (m, 2H), 6.56 (dt, J=8.6, 3.5 Hz, 1H), 6.50 (dd, J=6.0,2.9 Hz, 1H), 5.09 (d, J=1.3 Hz, 1H), 4.93 (d, J=1.3 Hz, 1H), 3.96 (d,J=3.6 Hz, 1H), 3.79 (s, 1H), 3.53 (s, 1H), 3.46 (s, 2H), 2.56-2.44 (m,1H), 2.28 (d, J=9.7 Hz, 1H), 2.10-2.02 (m, 2H), 1.96 (t, J=7.8 Hz, 2H),1.88-1.78 (m, 1H), 1.75 (dd, J=12.0, 6.4 Hz, 1H), 1.67 (dd, J=12.9, 5.8Hz, 1H), 1.35 (q, J=6.9 Hz, 1H), 1.32-1.15 (m, 6H), 0.86 (t, J=7.0 Hz,3H). ¹³C NMR (126 MHz, CDCl3) δ 154.69, 144.58, 135.54, 129.55, 127.85,127.61, 127.48, 126.70, 120.10, 119.69, 117.73, 115.55, 115.31, 114.90,82.02, 69.59, 55.42, 40.54, 33.61, 33.04, 31.66, 31.14, 30.02, 29.41,28.20, 27.26, 26.84, 25.35, 23.94, 23.51, 22.61, 14.09. 19F NMR (282MHz, CDCl3) δ −126.95 LRMS (ESI, APCI) m/z: calc'd for C28H37FNO [M+H]+421.3, found 420.9 FTIR (neat): 3361, 3085, 2956, 2930, 2856, 1494,1258, 1213, 775, 703, 668 cm-1.

(exo)-4-(5-amino-2-methylphenyl)-5-hexyl-3a-(1-phenylvinyl)-1,2,3,3a,6,6a-hexahydropentalen-1-ol(17g)

The general procedure for amine coupling was followed, using(exo)-4-(5-bromo-2-methylphenyl)-5-hexyl-3a-(1-phenylvinyl)-1,2,3,3a,6,6a-hexahydropentalen-1-olas the brominated [3.3.0] bicycle. The crude oil was purified by silicagel chromatography in 2030% EtOAc/Hex.

¹H NMR (500 MHz, Chloroform-d) δ 7.44-7.38 (m, 2H), 7.33-7.27 (m, 2H),7.02 (d, J=8.0 Hz, 1H), 6.85 (dd, J=8.9, 5.0 Hz, 1H), 6.54 (dd, J=8.0,2.5 Hz, 1H), 6.47 (d, J=2.5 Hz, 1H), 5.18 (d, J=1.1 Hz, 1H), 4.89 (d,J=1.2 Hz, 1H), 3.98 (s, 1H), 3.79 (s, 2H), 3.53 (s, 3H), 2.61 (dd,J=17.2, 10.0 Hz, 1H), 2.30 (d, J=9.7 Hz, 1H), 2.06-1.99 (m, 2H),1.97-1.79 (m, 2H), 1.77-1.68 (m, 2H), 1.38-1.07 (m, 11H), 0.86 (t, J=7.1Hz, 3H).

(exo)-4-(3-amino-S-fluorophenyl)-5-hexyl-3a-(1-phenylvinyl)-1,2,3,3a,6,6a-hexahydropentalen-1-ol(18g)

The general procedure for amine coupling was followed, using(exo)-4-(3-bromo-5-fluorophenyl)-5-hexyl-3a-(1-phenylvinyl)-1,2,3,3a,6,6a-hexahydropentalen-1-olas the brominated [3.3.0] bicycle. The crude oil was purified by silicagel chromatography in 20-30% EtOAc/Hex.

¹H NMR (500 MHz, CDCl₃) δ 7.36-7.30 (m, 2H), 7.31-7.20 (m, 3H),6.41-6.21 (m, 3H), 5.08 (d, J=1.6 Hz, 1H), 5.05 (d, J=1.6 Hz, 1H), 3.92(s, 1H), 3.79 (d, J=1.7 Hz, 1H), 3.70 (s, 2H), 3.53 (d, J=1.6 Hz, 1H),2.32 (dd, J=16.7, 9.3 Hz, 1H), 2.25 (d, J=9.4 Hz, 1H), 2.13-1.97 (m,4H), 1.79-1.63 (m, 3H), 1.42-1.12 (m, 6H), 0.87 (t, J=7.1, 6.5 Hz, 3H).¹³C NMR (126 MHz, CDCl3) δ 164.21, 162.28, 154.51, 147.23, 147.14,144.01, 141.78, 140.22, 140.14, 138.20, 127.73, 126.70, 119.67, 115.11,112.05, 106.87, 106.70, 100.65, 100.46, 82.00, 69.24, 55.89, 40.20,34.09, 31.98, 31.66, 31.14, 29.74, 29.40, 28.23, 27.74, 25.35, 23.91,23.52, 22.60, 14.08. ¹⁹F NMR (282 MHz, CDCl3) δ −114.23. LRMS (ESI,APCI) m/z: calc'd for C28H37FNO [M+H]+ 422.3, found 421.8 FTIR (neat):3360, 3210, 3085, 2956, 2929, 2858, 1692, 1610, 1585, 1459, 1425, 1258,756, 703 cm-1.

Synthetic Scheme for Modifications to External Styrene

General Procedure for Zirconecene Mediated Cyclization and MOMDeprotection.

Bis(cyclopentadienyl)zirconium(IV) dichloride (zirconecene dichloride)(1.2 equiv.) was dried by azeotroping away latent water with benzenefour times before being placed under nitrogen, dissolved in dry,degassed tetrahydrofuran (THF) and cooled to −78° C. in a dryice/acetone bath. The resulting solution of zirconecene dichloride wastreated with nBuLi (2.4 equiv.) to form a clear, light yellow solutionand allowed to stir. After approximately 30 minutes, azeotroped(5-(methoxymethoxy)hept-6-en-1-yn-1-yl)benzene (1.0 equiv.) in dry,degassed THF was added portionwise to afford a pink-orange solution, andthe reaction mixture was held at −78° C. for 30 minutes before allowingto warm to room temperature and stirred over 2.5 hours. The reactionmixture was then re-cooled to −78° C. and the required azeotropeddibromoheptane (1.1 equiv.) were added in dry, degassed THF. Freshlyprepared lithium diisopropylamine (LDA, 1.0 M, 1.1 equiv.) was added at−78° C. and stirred for 15 minutes. The required, freshly prepared,lithium acetylide (3.6 equiv.) was added to the reaction mixturedropwise in dry, degassed THF. The resulting dark reddish brown solutionwas stirred at −78° C. for 1.5 hours. The reaction was then quenchedwith methanol and saturated aqueous sodium bicarbonate and allowed towarm to room temperature to form a light yellow slurry. The slurry waspoured over water and extracted with ethyl acetate four times. Thecombined organic layers were washed with brine, dried with MgSO₄, andconcentrated in vacuo. The resulting colored oil (typically yellow,orange, or green), was roughly purified on a plug of silica and elutedwith 20% EtOAc/Hexanes to afford an oil which is a mixture of quenchedacetylide and desired bis-protected [3,3,0] bicyclic compounds (and insome cases protonolysis byproduct), which was carried on without furtherpurification.

The crude mixture (1.0 equiv.) was dissolved in acetonitrile, andconcentrated HCl (excess) was added and the resulting dark purple-bluereaction mixture was stirred for about 30 minutes (monitored by LCMS),concentrated, and subjected directly to silica gel chromatography in5-20% EtOAc/Hex to afford the desired exo compound.

(exo)-3a-(1-(2-fluorophenyl)vinyl)-5-hexyl-4-phenyl-1,2,3,3a,6,6a-hexahydropentalen-1-ol(20)

The general procedure for zirconecene mediated cyclization and MOMdeprotection was followed, using 1-ethynyl-2-fluorobenzene to form therequired lithium acetylide. The resulting crude oil was purified in5-20% EtOAc/Hex.

¹H NMR (500 MHz, CDCl₃) δ 7.38-7.17 (m, 7H), 7.03 (dtt, J=13.0, 7.5, 1.1Hz, 2H), 5.27 (s, 1H), 5.11 (s, 1H), 3.92 (d, J=4.3 Hz, 1H), 2.31 (d,J=14.3 Hz, 2H), 2.15-1.95 (m, 4H), 1.76-1.57 (m, 4H), 1.40-1.15 (m, 7H),0.86 (td, J=7.1, 0.9 Hz, 3H). ¹³C NMR (126 MHz, CDCl3) δ 160.57, 158.63,147.86, 141.76, 138.28, 137.40, 130.40, 129.67, 128.40, 128.34, 127.69,126.59, 123.18, 116.72, 115.43, 115.24, 81.97, 69.38, 56.83, 39.67,34.72, 31.65, 31.22, 29.77, 29.35, 27.91, 22.58, 14.08. ¹⁹F NMR (282MHz, CDCl3) δ −113.74. LRMS (ESI, APCI) m/z: calc'd for C28H36FO [M+H]+407.3, found 406.9 FTIR (neat): 3354, 3090, 3075, 2954, 2927, 2855,1489, 1446, 1217, 755, 701 cm-1.

(exo)-5-hexyl-4-phenyl-3a-(1-(o-tolyl)vinyl)-1,2,3,3a,6,6a-hexahydropentalen-1-ol(21)

The general procedure for zirconecene mediated cyclization and MOMdeprotection was followed, using 1-ethynyl-2-methylbenzene to form therequired lithium acetylide. The resulting crude oil was purified in 10%EtOAc/Hex.

¹H NMR (600 MHz, CDCl₃) δ 7.36-7.30 (m, 2H), 7.30-7.26 (m, 1H), 7.24 (d,J=7.5 Hz, 2H), 7.18 (d, J=7.5 Hz, 1H), 7.14 (t, J=7.4 Hz, 1H), 7.04 (t,J=7.4 Hz, 1H), 5.08 (s, 1H), 4.95 (s, 1H), 3.91 (s, 1H), 2.26 (s, 3H),2.25-2.12 (m, 2H), 2.06-1.88 (m, 4H), 1.76-1.66 (m, 2H), 1.61 (dd,J=11.9, 6.3 Hz, 1H), 1.42-1.28 (m, 3H), 1.28-1.22 (m, 1H), 1.22-1.15 (m,4H), 0.85 (t, J=7.2 Hz, 3H). ¹³C NMR (126 MHz, CDCl3) δ 152.27, 142.91,141.73, 138.48, 137.68, 135.64, 130.07, 129.97, 127.62, 126.62, 126.50,124.76, 115.47, 82.05, 74.70, 69.98, 55.74, 39.82, 34.59, 31.83, 31.67,29.65, 29.31, 27.91, 22.63, 20.74, 14.12. LRMS (ESI, APCI) m/z: calc'dfor C29H39O [M+H]+ 403.3, found 403.0 FTIR (neat): 3340, 3095, 3080,2954, 2926, 2855, 1489, 1456, 1440, 904, 765, 730, 710 cm-1.

(exo)-5-hexyl-3a-(1-(2-methoxyphenyl)vinyl)-4-phenyl-1,2,3,3a,6,6a-hexahydropentalen-1-ol(22)

The general procedure for zirconecene mediated cyclization and MOMdeprotection was followed, using 1-ethynyl-2-methoxylbenzene to form therequired lithium acetylide. The resulting crude oil was purified in10-20% EtOAc/Hex.

¹H NMR (600 MHz, CDCl₃) δ 7.36-7.18 (m, 6H), 6.95 (d, J=7.1 Hz, 1H),6.88-6.81 (m, 2H), 5.18 (d, J=1.8 Hz, 1H), 5.01 (d, J=1.8 Hz, 1H), 3.86(s, 1H), 3.75 (s, 3H), 2.50 (dd, J=16.6, 9.1 Hz, 1H), 2.45 (d, J=8.6 Hz,1H), 2.08 (d, J=16.6 Hz, 1H), 2.01 (t, J=7.7 Hz, 2H), 1.81-1.72 (m, 2H),1.65-1.58 (m, 2H), 1.58-1.53 (m, 1H), 1.36-1.29 (m, 2H), 1.27-1.20 (m,2H), 1.21-1.14 (m, 2H), 0.84 (t, J=7.2 Hz, 3H). ¹³C NMR (151 MHz, CDCl3)δ 172.46, 169.63, 156.37, 151.45, 140.47, 137.69, 132.61, 130.06,129.77, 127.97, 127.49, 126.39, 120.35, 115.58, 110.75, 81.67, 69.32,57.60, 55.56, 51.31, 39.63, 34.60, 31.63, 31.08, 29.60, 29.21, 28.00,22.58, 14.06. LRMS (ESI, APCI) m/z: calc'd for C29H39O2 [M+H]+ 417.3,found 417.9 FTIR (neat): 3355, 3085, 3075, 2950, 2928, 2855, 1738, 1490,1462, 1240, 1030, 751, 701 cm-1.

(exo)-3a-(1-(4-bromophenyl)vinyl)-5-hexyl-4-phenyl-1,2,3,3a,6,6a-hexahydropentalen-1-ol(23a)

The general procedure for zirconecene mediated cyclization and MOMdeprotection was followed, using 1-bromo-4-ethynylbenzene to form therequired lithium acetylide. The resulting crude oil was purified in 10%EtOAc/Hex.

(exo)-5-hexyl-3a-(1-(4-hydroxyphenyl)vinyl)-4-phenyl-1,2,3,3a,6,6a-hexahydropentalen-1-ol(23b)

Potassium Hydroxide (3.0 equiv.), tris(dibenzylideneacetone)dipalladium(0) (0.01 equiv.), and tBuXPhos (0.04 equiv.) were suspendedin 1,4-dioxane in a reaction tube under nitrogen.(exo)-3a-(1-(4-bromophenyl)vinyl)-5-hexyl-4-phenyl-1,2,3,3a,6,6a-hexahydropentalen-1-olwas added in dioxane. Water (˜10 equiv.) was added. The reaction mixturewas heated to 80° C. for 16 hours. After stirring, the mixture waspoured over water and ethyl acetate, and the organic layer was washedwith water and brine to remove 1,4-dioxane. The organic layer was driedwith MgSO₄, concentrated, and purified on silica in 20-50% EtOAc/Hex.

¹H NMR (500 MHz, CDCl₃) δ 7.38-7.27 (m, 4H), 7.24-7.14 (m, 3H),6.78-6.70 (m, 2H), 5.03 (d, J=1.4 Hz, 1H), 4.96 (d, J=1.5 Hz, 1H), 4.74(s, 1H), 3.95 (s, 1H), 2.38 (dd, J=16.9, 9.4 Hz, 1H), 2.28 (d, J=9.3 Hz,1H), 2.12-1.93 (m, 4H), 1.78-1.62 (m, 3H), 1.33 (dd, J=14.0, 6.7 Hz,3H), 1.31-1.17 (m, 5H), 0.86 (t, J=7.1 Hz, 3H). ¹³C NMR (126 MHz, CDCl3)δ 154.56, 153.99, 141.13, 139.08, 137.37, 136.62, 129.67, 128.96,127.61, 126.58, 114.52, 82.22, 69.46, 55.65, 40.27, 34.03, 32.03, 31.65,29.70, 29.38, 27.83, 22.60, 14.09. LRMS (ESI, APCI) m/z: calc'd forC28H37O2 [M+H]+ 404.3, found 403.9 FTIR (neat): 3323, 3095, 3070, 3040,2954, 2915, 2855, 1609, 1510, 1457, 1440, 1263, 1231, 837, 701 cm-1.

(exo)-5-hexyl-4-phenyl-3a-(3-phenylprop-1-en-2-yl)-1,2,3,3a,6,6a-hexahydropentalen-1-ol(24)

The general procedure for zirconecene mediated cyclization and MOMdeprotection was followed, using prop-2-yn-1-yl benzene to form therequired lithium acetylide. The resulting crude oil was purified in5-20% EtOAc/Hex. ¹H NMR (500 MHz, CDCl3) δ 7.34-7.27 (m, 4H), 7.25-7.17(m, 4H), 7.11-7.07 (m, 2H), 4.93 (d, J=1.1 Hz, 1H), 4.50 (d, J=1.4 Hz,1H), 3.99 (s, 1H), 3.50 (d, J=16.4 Hz, 1H), 3.39 (d, J=16.4 Hz, 1H),2.89 (dd, J=17.2, 9.3 Hz, 1H), 2.39 (d, 1H), 2.25 (dd, J=17.3, 2.0 Hz,1H), 2.17-2.03 (m, 3H), 1.80-1.70 (m, 1H), 1.56-1.51 (m, 1H), 1.42-1.35(m, 2H), 1.28-1.12 (m, 7H), 0.84 (t, J=7.1 Hz, 3H). ¹³C NMR (126 MHz,CDCl₃) δ 153.23, 140.85, 140.11, 139.13, 137.34, 129.65, 129.34, 128.28,127.66, 126.53, 125.96, 95.51, 82.47, 70.23, 55.58, 40.66, 39.43, 34.18,31.60, 30.21, 29.50, 29.18, 28.09, 22.61, 14.06. LRMS (ESI, APCI) m/z:calc'd for C29H39O [M+H]+ 403.3, found 403.0 FTIR (neat): 3362, 3070,3035, 2960, 2926, 2856, 1701, 1599, 1495, 1453, 1032, 908, 753, 732, 705cm-1.

(exo)-5-hexyl-1-hydroxy-4-phenyl-2,3,6,6a-tetrahydropentalen-3a(1H)-yl)(phenyl)methanone(25)

RJW100 (1.0 equiv.) was dissolved in DCM and cooled to −78° C. Ozone(excess) was bubbled through the solution until the reaction mixture wasblue. At this point, the stream of ozone was stopped and the reactionwas stirred until the blue color disappated. Dimethylsulfide (DMS) wasadded, and the reaction was briefly stirred. The reaction solution wasthen concentrated in vacuo and the crude reaction mixture was purifiedon silica in 0-20% EtOAc/Hex to afford a clear, colorless oil (14 mg,63%).

¹H NMR (600 MHz, CDCl₃) δ 7.86 (d, J=7.7 Hz, 2H), 7.46 (t, J=7.4 Hz,1H), 7.35 (dd, J=8.7, 6.7 Hz, 2H), 7.18 (d, J=6.6 Hz, 3H), 6.87 (dd,J=7.2, 2.3 Hz, 2H), 4.06 (s, 1H), 3.02 (dd, J=17.5, 10.2 Hz, 1H), 2.90(d, J=11.2 Hz, 1H), 2.76-2.64 (m, 1H), 2.33 (dd, J=17.7, 3.5 Hz, 1H),2.09 (t, J=7.8 Hz, 2H), 2.00 (d, J=12.2 Hz, 1H), 1.85-1.66 (m, 2H),1.49-1.36 (m, 2H), 1.29-1.15 (m, 6H), 0.85 (dd, J=14.0, 7.2 Hz, 3H). ¹³CNMR (101 MHz, CDCl3) δ 203.40, 141.99, 139.95, 138.62, 136.24, 131.77,129.03, 128.56, 128.13, 128.08, 126.99, 80.80, 76.25, 54.62, 40.48,32.65, 31.62, 30.50, 29.42, 29.28, 27.80, 22.58, 21.59, 14.07. LRMS(ESI, APCI) m/z: calc'd for C27H35O2 [M+H]+ 389.6, found 389.2 FTIR(neat): 3405, 3080, 2955, 2927, 2857, 1698, 1680, 1597, 1446, 1254,1180, 766, 699 cm-1.

(exo)-3a-benzyl-5-hexyl-4-phenyl-1,2,3,3a,6,6a-hexahydropentalen-1-ol(26)

A solution of 25 in ethylene glycol was prepared at room temperature.Hydrazine hydrate (9.5 equiv.) was added to the reaction mixture beforeheating to 100° C. for 1 h. Potassium hydroxide (10.0 equiv.) wassubsequently added and the reaction mixture was stirred at 150° C. for48 h. After stirring, the solution was cooled to room temperature,partitioned between water and EtOAc, and extracted with EtOAc 3×. Thecombined organic layers were dried with MgSO₄, filtered, andconcentrated in vacuo. The crude reaction mixture was purified on silicain 10-20% EtOAc/Hex (2.6 mg, 22%).

¹H NMR (600 MHz, CDCl₃) δ 7.37-7.33 (m, 3H), 7.30-7.27 (m, 1H),7.24-7.22 (m, 1H), 7.21-7.17 (m, 3H), 7.16-7.11 (m, 2H), 3.83 (s, 1H),2.93 (d, J=13.6 Hz, 1H), 2.73 (d, J=13.6 Hz, 1H), 2.38 (dd, J=15.6, 8.4Hz, 1H), 2.34 (d, J=10.6 Hz, 1H), 2.05-1.99 (m, 1H), 1.89-1.83 (m, 2H),1.74-1.67 (m, 3H), 1.49-1.44 (m, 1H), 1.29-1.18 (m, 5H), 1.12 (dt,J=6.0, 4.2 Hz, 3H), 0.82 (td, J=7.2, 0.6 Hz, 3H). ¹³C NMR (101 MHz,CDCl3) δ 140.91, 139.79, 139.35, 138.03, 130.45, 129.88, 127.91, 126.51,126.03, 81.83, 64.26, 52.58, 43.98, 38.94, 33.84, 32.06, 31.59, 29.22,29.02, 27.81, 22.56, 14.06. LRMS (ESI, APCI) m/z: calc'd for C26H35[M−H2O]+ 358.3, found 358.3 FTIR (neat): 3440, 3070, 3020, 2965, 2930,2855, 1498, 1456, 1263, 1185, 1029, 759, 703 cm-1.

Synthetic Scheme for Modifications to the Secondary Alcohol

1,1-Dibromoheptane

Triphenylphosphite (1.1 equiv) was suspended in DCM and cooled to −78°C. Bromine (1.1 equiv) was added dropwise and stirred briefly. Heptanal(1.0 equiv.) was then added in DCM and the reaction was allowed to cometo room temperature over 3 hours. The reaction mixture was then filteredthrough silica and concentrated in vacuo. The crude oil was purified bysilica gel chromatography in 100% hexanes to afford a clear, colorlessoil (67%).

RJW100 Synthesis General Procedure for Zirconecene Mediated Cyclizationand TBS Deprotection

Bis(cyclopentadienyl)zirconium(IV) dichloride (zirconecene dichloride)(1.2 equiv.) was dried by azeotroping away latent water with benzenefour times before being placed under nitrogen, dissolved intetrahydrofuran (THF) and cooled to −78° C. in a dry ice/acetone bath.The resulting solution of zirconecene dichloride was treated with nBuLi(2.4 equiv.) to form a clear, light yellow solution and allowed to stir.After approximately 30 minutes, azeotropedtert-butyldimethyl((7-phenylhept-1-en-6-yn-3-yl)oxy)silane (19) (1.0equiv.) in dry THE was added portionwise to afford a pink-orangesolution, and the reaction mixture was held at −78° C. for 30 minutesbefore allowing to warm and stir at room temperature for 2.5 hours. Thereaction mixture was then re-cooled to −78° C. and the requiredazeotroped dibromoheptane (1.1 equiv.) was added in dry THF. Freshlyprepared lithium diisopropylamine (LDA, 1.0 M, 1.1 equiv.) was added at−78° C. and stirred for 15 minutes. Lithium phenylacetylide (3.6 equiv.)was then prepared and added to the reaction mixture dropwise in dry THF.The resulting dark reddish brown solution was stirred at −78° C. for 1.5hours. The reaction was then quenched with methanol and saturatedaqueous sodium bicarbonate and allowed to warm to room temperature toform a light yellow slurry. The resulting slurry was poured over waterand extracted with ethyl acetate four times. The combined organic layerswere washed with brine, dried with MgSO₄, and concentrated in vacuo. Theresulting yellow oil was roughly purified on a plug of silica and elutedwith 20% EtOAc/Hexanes to afford a yellow oil which is a mixture ofphenylacetylene and desired bis-protected [3.3.0] bicyclic compounds,which was carried on without further purification. This procedureaffords exo and endo diastereomers in a 1.6:1 ratio as determined bycharacteristic 1H NMR signals.

The crude mixture (1.0 equiv.) was dissolved in THF, andtetrabutylammonium fluoride (1.5 equiv.) was added and the resultingdark brown solution was stirred open to air for 18 hours, concentrated,and subjected directly to silica gel chromatography in 5-10%EtOAc/Hexanes to separate both endo and exo diastereomers (with endoeluting before exo) of the desired compounds as clear, colorless oils.

5-hexyl-4-phenyl-3a-(1-phenylvinyl)-3,3a,6,6a-tetrahydropentalen-1(2H)-one(27)

A solution of alcohol (isomer irrelevant, 1.0 equiv.) in acetonitrilewas treated with N-methylmorpholine oxide (1.5 equiv.) and allowed tostir to homogeneity before the addition of tetrapropylammoniumperruthenate (0.1 equiv.) The solution was stirred at room temperatureuntil completion as determined by TLC (˜10 min). The solution wasconcentrated in vacuo and subjected directly to silica gelchromatography in 10% EtOAc/Hex to afford the title compound as a clear,colorless oil (88% yield).

¹H NMR (400 MHz, CDCl₃) δ 7.38-7.25 (m, 6H), 7.24-7.19 (m, 4H), 5.20 (d,J=1.4 Hz, 1H), 5.09 (d, J=1.4 Hz, 1H), 2.44 (d, J=7.5 Hz, 1H), 2.34-2.23(m, 2H), 2.15-1.95 (m, 5H), 1.89 (ddt, J=16.5, 7.8, 1.1 Hz, 1H),1.29-1.09 (m, 8H), 0.82 (t, J=7.0 Hz, 3H). ¹³C NMR (101 MHz, CDCl3) δ222.79, 153.19, 144.91, 142.47, 137.29, 136.63, 128.95, 128.24, 128.09,127.59, 127.03, 126.96, 115.26, 109.99, 65.39, 55.50, 38.76, 37.50,31.54, 29.97, 29.37, 28.33, 27.60, 22.52, 14.05. LRMS (ESI, APCI) m/z:calc'd for C28H35O [M+H]+ 385.3, found 385.3 FTIR (neat): 3080, 3053,2955, 2926, 2855, 1743, 1598, 1574, 1491, 1458, 1441, 906, 765, 701cm-1.

(endo orexo)-5-hexyl-4-phenyl-3a-(1-phenylvinyl)-1,2,3,3a,6,6a-hexahydropentalen-1-ylsulfamate (28 endo, 28 exo)

A 1M solution of sulfamoyl chloride (1.2 equiv.) in DMA was cooled to 0°C. A solution of the appropriate alcohol isomer (1.0 equiv.) in DMA wasadded slowly, followed by triethylamine (excess); the resulting solutionwas stirred for one hour. The solution was then diluted with water andextracted with EtOAc. The combined organic layers were then washed withwater and brine to remove DMA, dried with Na2SO4, filtered, andconcentrated in vacuo. The oil was purified by silica gel chromatographyin 20% EtOAc/Hex with 0.5% triethylamine, to afford the title compoundas a clear oil (endo, 78% yield).

Endo ¹H NMR (500 MHz, CDCl₃) δ 7.35-7.24 (m, 8H), 7.23-7.15 (m, 2H),5.11 (s, 1H), 4.92 (s, 1H), 4.87 (td, J=9.1, 5.2 Hz, 1H), 4.64 (s, 2H),2.71 (d, J=9.0 Hz, 1H), 2.60 (d, J=17.5 Hz, 1H), 2.17 (dd, J=17.7, 9.3Hz, 1H), 2.10-2.01 (m, 3H), 1.92-1.83 (m, 1H), 1.83-1.76 (m, 1H), 1.68(td, J=12.6, 5.6 Hz, 1H), 1.45-1.35 (m, 2H), 1.32-1.16 (m, 6H), 0.86 (t,J=7.1 Hz, 3H). Endo ¹³C NMR (126 MHz, CDCl3) δ 153.83, 143.49, 143.24,138.53, 136.48, 129.82, 127.88, 127.68, 127.62, 126.96, 126.78, 115.69,84.11, 68.22, 47.13, 34.89, 31.63, 31.15, 30.54, 29.76, 29.40, 27.73,22.59, 14.07. Endo LRMS (ESI, APCI) m/z: calc'd for C28H36NO3S [M−H]−465.3, found 465.4 Endo FTIR (neat): 3360, 3284, 3080, 3055, 3020, 2955,2928, 2855, 1558, 1491, 1441, 1356, 1184, 1029, 1004, 913, 851, 775,703, 668 cm-1.

Exo ¹H NMR (500 MHz, CDCl3) δ 7.36-7.28 (m, 7H), 7.29-7.16 (m, 3H), 5.10(d, J=1.3 Hz, 1H), 5.00 (d, J=1.3 Hz, 1H), 4.75 (d, J=4.4 Hz, 1H), 4.62(s, 2H), 2.68 (d, J=9.1 Hz, 1H), 2.40 (dd, J=18.1, 9.4 Hz, 1H),2.19-2.01 (m, 6H), 1.88-1.73 (m, 2H), 1.38-1.16 (m, 7H), 0.87 (t, J=7.1Hz, 3H). Exo ¹³C NMR (126 MHz, cdcl3) δ 153.55, 143.65, 141.37, 138.84,136.83, 129.56, 127.83, 127.77, 127.71, 126.89, 126.86, 115.59, 93.70,69.27, 52.76, 40.13, 32.14, 32.00, 31.60, 29.65, 29.36, 27.77, 22.56,14.06. Exo LRMS (ESI, APCI) m/z: calc'd for C28H36NO3S [M−H]− 465.3,found 465.2 Exo FTIR (neat): 3377, 3284, 3080, 3053, 3019, 2955, 2926,2854, 1598, 1572, 1558, 1491, 1457, 1440, 1356, 1182, 1073, 1028, 924,904, 813, 773, 764, 701, 667 cm-1.

(endo orexo)-5-hexyl-4-phenyl-3a-(1-phenylvinyl)-1,2,3,3a,6,6a-hexahydropentalen-1-ylcarbamate (29 endo, 29 exo)

The required alcohol isomer (1.0 equiv.) was suspended in MeCN andcooled to −15° C. Chlorosulfonyl isocyanate (2.0 equiv.) was added andthe reaction was stirred for 2 hours. After completion via TLC,concentrated hydrochloric acid (0.5) was added slowly and stirred toroom temperature over 4 hours. The solution was quenched with NaHCO₃,diluted with water, and extracted with EtOAc. The organic layers wererinsed with brine, dried with Na2SO4, filtered, and concentrated invacuo. The resulting oil was purified by silica gel chromatography in5-30% EtOAc/Hex to afford the title compounds as yellow oils (79%).

Endo ¹H NMR (400 MHz, CDCl₃) δ 7.34-7.23 (m, 5H), 7.24-7.18 (m, 5H),5.04 (d, J=1.4 Hz, 1H), 4.93 (d, J=1.4 Hz, 1H), 4.91-4.87 (m, OH), 4.54(s, 2H), 2.66 (td, J=8.9, 1.8 Hz, 1H), 2.32 (dd, J=17.7, 1.9 Hz, 1H),2.10-1.95 (m, 3H), 1.93-1.83 (m, 1H), 1.73-1.59 (m, 2H), 1.36 (p, J=7.3Hz, 2H), 1.29-1.16 (m, 7H), 0.84 (t, J=7.0 Hz, 3H). Endo ¹³C NMR (101MHz, CDCl₃) δ 156.40, 154.31, 143.67, 143.27, 138.53, 136.90, 129.71,127.77, 127.69, 127.63, 126.73, 126.59, 115.16, 68.54, 46.99, 34.45,31.70, 31.07, 30.10, 29.79, 29.39, 27.81, 22.61, 14.10. Endo LRMS (ESI,APCI) m/z: calc'd for C29H39NO3 [M+H2O]− 449.3, 449.1 Endo FTIR (neat):3490, 3343, 3085, 3050, 3015, 2955, 2926, 2855, 1716, 1598, 1491, 1440,1392, 1335, 1041, 903, 765, 701 cm-1.

Exo ¹H NMR (500 MHz, CDCl3) δ 7.41-7.19 (m, 10H), 5.08 (d, J=1.6 Hz,1H), 5.02 (d, J=1.6 Hz, 1H), 4.77 (dt, J=4.2, 1.4 Hz, 1H), 4.56 (s, 2H),2.42 (d, J=8.1 Hz, 1H), 2.36 (dd, J=16.3, 8.9 Hz, 1H), 2.19 (d, J=17.0Hz, 1H), 2.14-1.88 (m, 4H), 1.85-1.63 (m, 3H), 1.41-1.29 (m, 2H),1.31-1.16 (m, 5H), 0.88 (t, J=7.0 Hz, 3H). Exo ¹³C NMR (126 MHz, CDCl3)δ 156.59, 154.47, 143.89, 141.92, 138.53, 137.30, 129.61, 127.76,127.71, 126.75, 126.67, 115.02, 85.61, 69.36, 53.01, 40.34, 32.36,31.66, 31.54, 29.74, 29.40, 27.81, 22.60, 14.11. Exo LRMS (ESI, APCI)m/z: calc'd for C29H39NO3 [M+H2O]− 449.3, 449.3 Exo FTIR (neat): 3350,3194, 3055, 2956, 2927, 2855, 1712, 1597, 1544, 1492, 1443, 1408, 1336,1077, 819, 759, 702 cm-1.

(endo or exo)5-hexyl-4-phenyl-3a-(1-phenylvinyl)-1,2,3,3a,6,6a-hexahydropentalen-1-ylmethanesulfonate (30 endo 30 exo)

In a 20 dram vial equipped with a stir bar was dissolved the desiredisomer of RJW100 (1.0 equiv.). Triethylamine (5.0 equiv.) was added,followed by methanesulfonyl chloride (5.0 equiv.) The reaction mixturewas allowed to stir at room temperature for 1 h before concentrating invacuo and purifying by silica gel chromatography in 30% EtOAc/hexanes.(Endo: 62.1 mg, 95%; Exo: 150.3 mg, 95%)

Endo ¹H NMR (500 MHz, CDCl₃) δ 7.36-7.26 (m, 8H), 7.24-7.17 (m, 2H),5.13 (d, J=1.1 Hz, 1H), 5.04-4.92 (m, 1H), 4.95 (d, J=1.2 Hz, 1H), 3.00(s, 3H), 2.70 (t, J=9.0, 1.8 Hz, 1H), 2.60 (d, J=17.4 Hz, 1H), 2.17 (dd,J=17.5, 9.1 Hz, 1H), 2.08 (ttd, J=13.5, 6.8, 4.9 Hz, 4H), 1.92-1.76 (m,3H), 1.72 (td, J=12.5, 5.8 Hz, 1H), 1.40 (p, 2H), 1.33-1.18 (m, 4H),0.88 (t, J=7.2 Hz, 3H). Endo ¹³C NMR (126 MHz, CDCl3) δ 153.71, 143.41,143.19, 138.52, 136.45, 129.78, 127.89, 127.72, 127.64, 126.99, 126.81,115.69, 82.85, 68.23, 47.40, 38.23, 34.86, 31.65, 31.11, 30.97, 29.76,29.41, 27.76, 22.60, 14.09. Endo LRMS (ESI, APCI) m/z: calc'd forC29H38O3S [M] 466.3, found [M-CH3O3S] 368.9 Endo FTIR (neat): 3070,3028, 2955, 2931, 2857, 1570, 1492, 1445, 1356, 1180, 938, 908, 857,764, 702 cm-1.

Exo ¹H NMR (500 MHz, CDCl3) δ 7.37-7.25 (m, 8H), 7.27-7.19 (m, 2H), 5.11(d, J=1.3 Hz, 1H), 5.01 (d, J=1.3 Hz, 1H), 4.83 (d, J=4.0 Hz, 1H), 2.95(s, 3H), 2.63 (d, J=9.2 Hz, 1H), 2.41 (dd, J=17.4, 9.5 Hz, 1H), 2.14(dd, J=17.5, 2.0 Hz, 1H), 2.11-1.98 (m, 4H), 1.90-1.75 (m, 2H),1.40-1.31 (m, 2H), 1.32-1.17 (m, 6H), 0.87 (t, J=7.1 Hz, 3H). Exo ¹³CNMR (151 MHz, CDCl3) δ 153.50, 143.57, 141.32, 138.78, 136.75, 132.75,129.55, 127.83, 127.76, 127.64, 126.90, 126.86, 115.62, 92.12, 69.21,53.04, 39.96, 38.73, 32.36, 32.11, 31.58, 29.62, 29.35, 27.75, 22.55,14.05. Exo LRMS (ESI, APCI) m/z: calc'd for C29H38O3S [M] 466.3, found[M-CH3O3S] 368.9 Exo FTIR (neat): 3060, 3028, 2960, 2929, 2855, 1560,1544, 1498, 1441, 1354, 1173, 930, 901, 702 cm-1.

(endo orexo)-5-hexyl-4-phenyl-3a-(1-phenylvinyl)-1,2,3,3a,6,6a-hexahydropentalen-1-azide(31 endo, 31 exo)

In a reaction tube equipped with a stir bar was added the requiredmesylated isomer (30 endo or exo, 1.0 equiv.) in DMF. Sodium azide (10.0equiv.) was added and the reaction mixture was allowed to stir for about16 h at 80° C. After stirring, the solution was allowed to cool to roomtemperature and poured over water and EtOAc. The organic layer waswashed with water and brine in order to remove DMF, dried over MgSO₄,and concentrated in vacuo. The reaction mixture was purified on silicain 0-10% EtOAc/hex. (Endo: 117.2 mg, 88%; Exo: 45.6 mg, 90%) (Note:inversion of stereochemistry).

Endo ¹H NMR (600 MHz, CDCl₃) δ 7.36-7.26 (m, 8H), 7.23-7.18 (m, 2H),5.10 (d, J=1.3 Hz, 1H), 4.94 (d, J=1.3 Hz, 1H), 3.87 (ddd, J=10.5, 8.8,5.9 Hz, 1H), 2.62-2.51 (m, 2H), 2.16-2.01 (m, 4H), 1.97-1.88 (m, 1H),1.79 (ddd, J=12.4, 5.9, 1.8 Hz, 1H), 1.71 (td, J=12.4, 5.2 Hz, 1H),1.67-1.59 (m, 1H), 1.40 (p, J=7.5 Hz, 2H), 1.31-1.19 (m, 5H), 0.87 (t,J=7.2 Hz, 3H). Endo ¹³C NMR (126 MHz, CDCl3) δ 154.20, 143.75, 143.36,138.51, 136.77, 129.82, 127.84, 127.65, 126.89, 126.70, 115.50, 69.05,64.86, 47.89, 35.65, 32.51, 31.68, 30.15, 29.80, 29.45, 27.77, 22.62,14.12. Endo LRMS [ESI] calc'd for C₂₈H₃₄N₃ 412.3 [M+H]+, found 411.8Endo FTIR (neat): 3085, 3053, 2955, 2929, 2855, 2098 (s), 1491, 1441,1340, 1259, 905, 775, 763, 701 cm-1.

Exo ¹H NMR (500 MHz, CDCl3) δ 7.36-7.24 (m, 8H), 7.21 (dt, J=7.5, 1.4Hz, 2H), 5.09 (s, 1H), 5.00 (s, 1H), 3.64 (s, 1H), 2.44-2.35 (m, 2H),2.14-1.93 (m, 5H), 1.83-1.67 (m, 3H), 1.40-1.30 (m, 2H), 1.32-1.17 (m,5H), 0.86 (d, J=7.1 Hz, 3H). Exo ¹³C NMR (126 MHz, CDCl3) δ 153.83,143.76, 141.32, 139.07, 137.03, 129.64, 127.78, 127.77, 127.72, 126.80,126.75, 115.29, 71.33, 69.34, 52.09, 41.13, 32.64, 31.63, 31.20, 29.69,29.38, 27.80, 22.58, 14.07. Exo LRMS (ESI) m/z: calc'd for C₂₈H₃₄N₃[M+H]+ 412.3, found 412.3 Exo FTIR (neat): 3080, 3053, 3019, 2955, 2926,2855, 2097 (s), 1491, 1441, 1341, 1247, 904, 774, 701 cm-1.

(endo orexo)-5-hexyl-4-phenyl-3a-(1-phenylvinyl)-1,2,3,3a,6,6a-hexahydropentalene-1-carbonitrile(32 endo, 32 exo)

Sodium cyanide (10.0 equiv.) was suspended in DMF, followed by additionof the required mesylated isomer (30 endo or 30 exo) (1.0 equiv.) inDMF. The mixture was allowed to stir at 100° C. for about 40 hours. Thereaction was diluted with water and extracted with EtOAc. The combinedorganic layers were washed with water and brine to remove DMF, driedwith Na2SO4, filtered, and concentrated to dryness. The crude oil waspurified by silica gel chromatography in 5% EtOAc/Hex to yield the titlecompound. (Endo: 21.9 mg, 26%; Exo: 19.9 mg, 47%) (Note: An appreciableamount of E2 elimination product is typically also observed, despiteconsiderable optimization of reaction conditions. Also note theinversion of stereochemistry).

Endo ¹H NMR (600 MHz, CDCl₃) δ 7.34-7.23 (m, 8H), 7.22-7.19 (m, 2H),5.08 (s, 1H), 4.98 (s, 1H), 2.91-2.84 (m, 1H), 2.60 (t, J=9.0 Hz, 1H),2.53 (d, J=17.5 Hz, 1H), 2.30 (dd, J=17.6, 8.6 Hz, 1H), 2.15-1.97 (m,3H), 1.83-1.75 (m, 2H), 1.74-1.66 (m, 1H), 1.39 (p, J=7.6 Hz, 2H),1.31-1.17 (m, 6H), 0.84 (d, J=7.0 Hz, 3H). Endo ¹³C NMR (126 MHz, CDCl3)δ 153.37, 143.23, 143.22, 137.89, 136.55, 129.57, 127.88, 127.83,127.73, 127.05, 126.90, 121.12, 115.80, 69.77, 46.49, 39.10, 34.87,34.69, 31.59, 30.57, 29.75, 29.46, 27.68, 22.59, 14.07. Endo LRMS (ESI,APCI) m/z: calc'd for C29H35N [M+H]+ 396.6 found 396.4 Endo FTIR (neat):3075, 3053, 3025, 2950, 2926, 2854, 2237, 1598, 1573, 1491, 1442, 1378,1074, 1029, 907, 765, 702 cm-1.

Exo ¹H NMR (600 MHz, CDCl3) δ 7.38-7.20 (m, 10H), 5.09 (s, 1H), 5.08 (s,1H), 2.71 (dd, J=7.9, 4.9 Hz, 1H), 2.53 (q, J=11.0, 5.9 Hz, 1H), 2.29(dd, J=17.9, 8.4 Hz, 1H), 2.12-1.98 (m, 4H), 1.92-1.86 (m, 2H), 1.72(dt, J=13.1, 5.2 Hz, 1H), 1.36-1.15 (m, 8H), 0.84 (t, J=7.0 Hz, 3H). Exo¹³C NMR (75 MHz, CDCl3) δ 153.05, 142.91, 141.48, 138.61, 136.60,129.40, 128.01, 127.92, 127.82, 126.99, 126.95, 123.09, 115.43, 69.57,51.76, 41.68, 37.48, 33.78, 31.60, 30.52, 29.74, 29.41, 27.79, 22.57,14.08. Exo LRMS (ESI, APCI) m/z: calc'd for C29H36N [M+H]+ 398.3, found398.3 Exo FTIR (neat): 3080, 3051, 3020, 2950, 2926, 2854, 2236, 1598,1573, 1491, 1441, 1378, 1074, 1029, 905, 768, 700 cm-1.

(endo orexo)-5-hexyl-4-phenyl-3a-(1-phenylvinyl)-1,2,3,3a,6,6a-hexahydropentalen-1-amine(33 endo, 33 exo)

The required azide isomer (31 endo or 31 exo) (1.0 equiv.) was dissolvedin anhydrous Et2O under nitrogen then LiAlH4 (4.0M in Et2O, 10.0 equiv.)was added dropwise and stirred at room temperature for ˜1 hr, until thereaction was complete by TLC. The reaction was quenched by cooling to 0°C. and diluting with Et2O before slow addition of 1 mL per gram ofLiAlH4 water then 4 M NaOH. before pouring over EtOAc. The organic layerwas washed with Rochelle's salt and brine to remove DMF and lithiumsalts, dried over MgSO4, and concentrated in vacuo. The resulting oilwas then purified by silica gel chromatography in 50% EtOAc/Hex with 1%triethylamine to afford a colorless oil. (endo: 47.9 mg, 95%, exo: 40.0mg, 92%).

Endo ¹H NMR (600 MHz, CDCl3) δ 7.37-7.19 (m, 10H), 5.08 (d, J=1.4 Hz,1H), 4.94 (d, J=1.5 Hz, 1H), 3.30 (ddd, J=11.0, 8.8, 5.7 Hz, 1H), 2.48(d, J=17.4 Hz, 1H), 2.42 (t, J=9.0 Hz, 1H), 2.12-2.00 (m, 2H), 1.83-1.78(m, 1H), 1.73-1.68 (m, 2H), 1.46-1.37 (m, 2H), 1.35-1.20 (m, 8H), 0.88(t, J=7.1 Hz, 3H). Endo ¹³C NMR (151 MHz, CDCl3 δ 155.08, 144.23,142.88, 139.44, 137.15, 129.78, 127.72, 127.66, 127.56, 126.61, 126.49,115.01, 69.46, 55.31, 49.06, 34.55, 34.08, 33.25, 31.67, 29.87, 29.49,27.97, 22.63, 14.11. Endo LRMS (ESI, APCI) m/z: calc'd for C28H36N[M+H]+ 386.28, found 385.9 Endo FTIR 3079, 3052, 3052, 3018, 2951, 2926,2854, 1667, 1614, 1598, 1572, 1490, 1458, 1441, 1378, 1330, 1264, 1239,1180, 1155, 1102, 1074, 1029, 1001, 945, 903, 845, 775, 761, 738, 700,677 cm-1.

Exo ¹H NMR (600 MHz, CDCl3) δ 7.33-7.28 (m, 4H), 7.28-7.21 (m, 6H), 5.04(d, J=1.5 Hz, 1H), 5.03 (d, J=1.4 Hz, 1H), 3.01 (dt, J=5.4, 3.9 Hz, 1H),2.32-2.26 (m, 1H), 2.08-2.02 (m, 4H), 1.79-1.72 (m, 1H), 1.65-1.60 (m,1H), 1.42-1.17 (m, 10H), 0.85 (t, J=7.2 Hz, 3H). Exo ¹³C NMR (126 MHz,CDCl3) δ 153.82, 143.42, 141.30, 139.03, 137.13, 129.50, 128.11, 127.77,127.63, 126.71, 126.67, 114.91, 69.20, 61.18, 40.53, 32.10, 31.62,30.31, 29.69, 29.43, 27.82, 22.57, 14.06. Exo LRMS (ESI, APCI) m/z:calc'd for C28H36N [M+H]+ 386.28, found 385.9 Exo FTIR 3079, 3052, 3018,2953, 2926, 2855, 1618, 1597, 1572, 1491, 1458, 1440, 1378, 1331, 1264,1181, 1155, 1074, 1028, 962, 901, 844, 803, 764, 736, 700, 679 cm-1.

(endo or exo)5-hexyl-4-phenyl-3a-(1-phenylvinyl)-1,2,3,3a,6,6a-hexahydropentalen-1-yl)-1H-1,2,3-triazole(34 endo, 34 exo)

In a 20 dram vial, ascorbic acid (1.0 equiv.) and potassium carbonate(6.0 equiv.) were dissolved in water. Copper sulfate pentahydrate (1.0equiv.) was then added and the reaction was stirred briefly.Trimethysilyl acetylene (6.0 equiv.) was added in MeOH and the crudemixture of required azide isomer (31 endo or 31 exo) (1.0 equiv.) wassubsequently added. The reaction mixture was allowed to stir overnightbefore dilution with water and extraction with EtOAc. The combinedorganics were washed with brine and dried over MgSO₄ beforeconcentration in vacuo. The crude oil was purified by silica gelchromatography in 30% EtOAc/hexanes.

Endo ¹H NMR (600 MHz, CDCl₃) δ 7.70 (s, 1H), 7.49 (s, 1H), 7.36-7.25 (m,8H), 7.24-7.21 (m, 2H), 5.14 (s, 1H), 5.00 (s, 1H), 4.96 (ddd, J=11.5,9.5, 6.7 Hz, 1H), 2.94 (td, J=9.2, 1.9 Hz, 1H), 2.29-2.20 (m, 2H),2.03-1.84 (m, 5H), 1.27-1.09 (m, 9H), 0.81 (t, J=7.0 Hz, 3H). Endo ¹³CNMR (126 MHz, CDCl3) δ 153.77, 143.46, 138.44, 133.33, 129.70, 127.91,127.83, 127.69, 127.03, 126.93, 122.88, 115.86, 69.13, 63.41, 48.60,35.66, 32.60, 31.54, 29.74, 29.39, 29.34, 27.65, 22.55, 14.06. Endo LRMS(ESI, APCI) m/z: calc'd for C₃₀H₃₈N₃ [M+H]+ 440.3, found 440.4 Endo FTIR(neat): 3080, 3053, 2956, 2927, 2854, 1598, 1573, 1491, 1441, 1288,1073, 1029, 905, 775, 702 cm-1.

Exo ¹H NMR (600 MHz, CDCl₃) δ 7.66 (s, 1H), 7.43 (s, 1H), 7.37-7.20 (m,10H), 5.13 (dd, J=0.9 Hz, 1H), 5.09 (d, J=0.9 Hz, 1H), 4.79-4.72 (m,1H), 2.79 (dd, J=8.6, 3.8 Hz, 1H), 2.41 (dd, J=17.2, 8.6 Hz, 1H), 2.28(d, J=17.5 Hz, 1H), 2.18-1.98 (m, 4H), 1.85-1.77 (m, 1H), 1.40-1.33 (m,2H), 1.29-1.16 (m, 7H), 0.85 (t, J=7.1 Hz, 3H). Exo ¹³C NMR (101 MHz,CDCl3) δ 153.39, 143.17, 141.25, 139.19, 136.72, 129.56, 127.96, 127.87,127.77, 127.08, 126.92, 121.73, 115.68, 109.99, 69.99, 69.30, 53.10,41.18, 32.97, 32.88, 31.61, 29.75, 29.41, 27.82, 22.58, 14.08. Exo LRMS(ESI, APCI) m/z: calc'd for C₃₀H₃₈N₃ [M+H]+ 4.3, found 439.3 Exo FTIR(neat): 3088, 3055, 2955, 2925, 2855, 1599, 1491, 1446, 1265, 1074, 910,773, 703 cm-1.

N-((endo orexo)-5-hexyl-4-phenyl-3a-(1-phenylvinyl)-1,2,3,3a,6,6a-hexahydropentalen-1-yl)acetamide(35 endo, 35 exo)

A DCM solution of acetyl chloride (1.5 equiv.) was cooled to 0° C., andslowly treated with a solution of the required amine isomer (33 endo or33 exo) (1.0 equiv.) in DCM, followed by triethylamine (3.0 equiv.); theresulting solution was stirred for one hour. The solution was thendiluted with water and extracted with DCM. The combined organic layerswere then washed with water and brine, dried with Na2SO4, filtered, andconcentrated in vacuo. The oil was then purified by silica gelchromatography in 35% EtOAc/Hex to afford the title compound as a yellowoil. (71%)

Endo ¹H NMR (500 MHz, CDCl₃) δ 7.35-7.27 (m, 5H), 7.25-7.22 (m, 5H),5.35 (d, J=8.1 Hz, 1H), 5.06 (d, J=1.5 Hz, 1H), 5.02 (d, J=1.5 Hz, 1H),4.25 (dtd, J=10.5, 8.6, 6.2 Hz, 1H), 2.66 (ddd, J=16.9, 8.4, 1.6 Hz,1H), 2.14-2.00 (m, 4H), 1.99 (s, 3H), 1.87 (dtd, J=11.7, 6.0, 2.3 Hz,1H), 1.76 (td, J=12.2, 11.7, 5.8 Hz, 1H), 1.66 (ddd, J=12.7, 5.9, 2.3Hz, 1H), 1.43-1.26 (m, 1H), 1.30-1.18 (m, 8H), 0.87 (t, J=7.0 Hz, 3H).Endo ¹³C NMR (126 MHz, CDCl3) δ 169.34, 154.62, 143.56, 141.66, 138.87,137.23, 129.62, 127.87, 127.78, 127.74, 126.87, 126.64, 114.82, 68.95,59.48, 54.41, 40.86, 32.98, 32.10, 31.64, 29.80, 29.42, 27.82, 25.99,23.56, 22.59, 14.08. Endo LRMS (ESI, APCI) m/z: calc'd for C30H39NO[M+H]+ 430.3, found 430.3 Endo FT-IR (neat): 3284, 3079, 3055, 2954,2924, 2853, 1646, 1551, 1491, 1441, 1375, 1301, 1249, 902, 774, 701, 668cm-1.

Exo ¹H NMR (500 MHz, CDCl₃) δ 7.35-7.25 (m, 8H), 7.27-7.19 (m, 2H), 5.35(d, J=7.4 Hz, 1H), 5.07 (s, 2H), 3.96 (dd, J=8.2, 4.0 Hz, 1H), 2.34 (dd,J=17.2, 8.6 Hz, 1H), 2.25 (d, J=7.1 Hz, 1H), 2.22-2.16 (m, 1H),2.08-2.02 (m, 2H), 1.93 (s, 3H), 1.90-1.82 (m, 2H), 1.75-1.66 (m, 1H),1.40-1.14 (m, 9H), 0.86 (t, J=7.1 Hz, 3H). Exo ¹³C NMR (126 MHz, CDCl3)δ 169.45, 154.32, 143.48, 143.00, 141.25, 139.28, 136.88, 129.47,127.99, 127.79, 127.65, 126.72, 114.96, 69.04, 53.14, 47.38, 35.24,32.05, 31.68, 29.90, 29.55, 28.13, 23.34, 22.62, 14.09. Exo LRMS (ESI,APCI) m/z: calc'd for C30H39NO [M+H]+ 430.3, found 430.3 Exo FTIR(neat): 3280, 3079, 3055, 2955, 2854, 1646, 1598, 1550, 1491, 1441,1375, 1304, 1178, 1074, 1029, 903, 766, 701, 668 cm-1.

1-((endo orexo)-5-hexyl-4-phenyl-3a-(1-phenylvinyl)-1,2,3,3a,6,6a-hexahydropentalen-1-yl)urea(36 endo, 36 exo)

To a reaction flask charged with the required amine isomer (33 endo or33 exo) (1.0 equiv.) and sodium cyanate (10.0 equiv.) was added waterand 1M aqueous hydrochloric acid (2.0 equiv.). The reaction mixture washeated to 90° C. and stirred for approximately 72 hours at thistemperature before being diluted with 3M aqueous NaOH and extracted withEt2O. The combined organic layers were then washed with water and brine,dried with Na2SO4, filtered, and concentrated in vacuo. The oil was thenpurified by silica gel chromatography in EtOAc to afford the titlecompound as a white solid. (Endo: 6.9 mg, 41%; Exo: 4.5 mg, 37%).

Endo ¹H NMR (600 MHz, CDCl3) δ 7.34-7.19 (m, 10OH), 5.06 (d, J=1.4 Hz,1H), 4.99 (d, J=1.4 Hz, 1H), 4.48 (d, J=7.7 Hz, 1H), 4.32 (s, 2H), 3.97(s, 1H), 2.60 (t, J=8.7 Hz, 1H), 2.22 (d, J=17.3 Hz, 1H), 2.09-2.03 (m,2H), 1.93-1.85 (m, 1H), 1.73 (td, J=12.5, 5.7 Hz, 1H), 1.67 (ddd,J=12.9, 6.1, 1.9 Hz, 1H), 1.39-1.31 (m, 2H), 1.29-1.17 (m, 8H), 0.86 (t,J=7.1 Hz, 3H). Endo 13C NMR (126 MHz, CDCl3) δ 157.89, 154.39, 143.57,143.15, 139.01, 136.83, 129.54, 127.88, 127.72, 127.68, 126.76, 126.69,115.11, 69.16, 47.58, 35.05, 32.13, 31.87, 31.63, 29.89, 29.51, 27.97,22.60, 14.07. Endo LRMS (ESI, APCI) m/z: calc'd for C29H37N2O [M+H]+429.7, found 428.9 Endo FT-IR (neat): 3348, 3080, 3053, 3018, 2952,2923, 2853, 1740, 1655, 1599, 1552, 1491, 1458, 1377, 1341, 1287, 1234,1212, 1156, 1104, 1075, 1029, 966, 902, 860, 773, 763, 725, 701, 669cm-1.

Exo ¹H NMR (600 MHz, CDCl3) δ 7.34-7.19 (m, 10H), 5.06 (d, J=1.3 Hz,1H), 5.03 (d, J=1.4 Hz, 1H), 4.42 (d, J=7.4 Hz, 1H), 4.25 (s, 2H), 3.70(s, 1H), 2.36 (dd, J=17.2, 8.9 Hz, 1H), 2.22-2.17 (m, 2H), 2.05 (q,J=7.0 Hz, 2H), 1.94-1.80 (m, 2H), 1.72-1.66 (m, 1H), 1.59-1.52 (m, 1H),1.33 (q, J=7.4 Hz, 2H), 1.28-1.17 (m, 6H), 0.85 (t, J=7.2 Hz, 3H). Exo13C NMR (126 MHz, CDCl3) δ 157.74, 154.49, 143.59, 141.50, 138.97,137.17, 129.61, 127.83, 127.77, 127.71, 126.84, 126.65, 114.89, 68.96,60.87, 54.41, 41.05, 32.81, 32.37, 31.62, 29.76, 29.69, 29.40, 27.81,22.57, 14.06. Exo LRMS (ESI, APCI) m/z: calc'd for C₂₉H₃₇N₂O [M+H]+429.7, found 428.9 Exo FTIR (neat): 3317, 3079, 3053, 3018, 2954, 2923,2854, 1641, 1591, 1545, 1491, 1459, 1440, 1378, 1339, 1261, 1195, 1182,1157, 1075, 1028, 903, 844, 803, 775, 764, 720, 700, 669 cm-1.

(endo orexo)-5-hexyl-4-phenyl-3a-(1-phenylvinyl)-1,2,3,3a,6,6a-hexahydropentalen-1-ylsulfamide (37 endo or 37 exo)

To a solution of the required amine isomer (33 endo or 33 exo) (1.1equiv.) and triethylamine (2.0 equiv.) in DCM under nitrogen was added a0.5M solution of 2-oxo-1,3-oxazolidine-3-sulfonyl chloride in DCM (1.0equiv.) (prepared according to the procedure of Borghese et al.). Theresulting solution was stirred at room temperature for three hours thenconcentrated in vacuo to a solid. To the resulting crude solid was addedammonia (0.5M in dioxane, 1.5 equiv.) and triethylamine (3.0 equiv.).The solution was then heated in a sealed tube to 85° C. overnight. Aftercooling to room temperature, the reaction was diluted with 3:3:94MeOH:Et3N:EtOAc then passed through a pad of silica. The eluent wasconcentrated in vacuo and purified by silica gel chromatography in20-30% EtOAc/hexanes to afford the title compound as a colorless oil.(Endo: 21.6 mg, 60%; Exo: 5.4 mg, 36%)

Endo ¹H NMR (600 MHz, CDCl₃) δ 7.33-7.23 (m, 8H), 7.20-7.17 (m, 2H),5.09 (d, J=1.3 Hz, 1H), 4.96 (d, J=1.3 Hz, 1H), 4.44 (s, 2H), 4.36 (d,J=8.0 Hz, 1H), 3.84-3.77 (m, 1H), 2.62 (td, J=8.9, 2.0 Hz, 1H), 2.38(dd, J=17.5, 2.0 Hz, 1H), 2.20-2.13 (m, 1H), 2.08-2.04 (m, 2H),2.00-1.95 (m, 1H), 1.74-1.70 (m, 2H), 1.50-1.43 (m, 1H), 1.42-1.16 (m,8H), 0.86 (t, J=7.1 Hz, 3H). Endo ¹³C NMR (126 MHz, CDCl3) δ 154.13,143.56, 142.84, 139.30, 136.58, 129.64, 127.80, 127.74, 126.87, 126.78,115.49, 68.80, 57.16, 47.44, 35.42, 32.32, 31.97, 31.60, 29.83, 29.48,27.92, 22.59, 14.07. Endo LRMS (ESI, APCI) m/z: calc'd for C28H37N2O2S[M+H]+ 465.7, found 464.8 Endo FT-IR (neat): 3278, 3080, 3053, 3019,2954, 2926, 2854, 1718, 1618, 1598, 1571, 1491, 1440, 1323, 1160, 1118,1095, 1075, 1029, 1014, 906, 774, 763, 720, 700 cm-1.

Exo ¹H NMR (600 MHz, CDCl3) δ 7.34-7.24 (m, 8H), 7.23-7.18 (m, 2H), 5.08(d, J=1.2 Hz, 1H), 5.01 (d, J=1.2 Hz, 1H), 4.38 (s, 2H), 4.21 (d, J=7.2Hz, 1H), 3.58-3.52 (m, 1H), 2.40 (dd, J=16.9, 8.9 Hz, 1H), 2.36-2.31 (m,1H), 2.18 (d, J=16.9 Hz, 1H), 2.05 (td, J=7.5, 2.6 Hz, 2H), 1.99-1.85(m, 2H), 1.76-1.69 (m, 2H), 1.38-1.30 (m, 1H), 1.30-1.15 (m, 7H), 0.86(t, J=7.2 Hz, 3H). Exo ¹³C NMR (126 MHz, CDCl3) δ 154.08, 143.53,141.15, 139.27, 136.94, 129.62, 127.86, 127.74, 126.93, 126.75, 115.21,68.77, 63.78, 54.03, 40.83, 32.84, 32.30, 31.62, 29.71, 29.69, 29.37,27.81, 22.57, 14.05. Exo LRMS (ESI, APCI) m/z: calc'd for C₂₉H₃₇N₂O[M+H]+ 465.7, found 464.8 Exo FTIR (neat): 3277, 3080, 3053, 3018, 2923,2854, 1720, 1621, 1598, 1572, 1491, 1455, 1441, 1376, 1321, 1158, 1075,1029, 969, 903, 765, 723, 701, 668 cm-1.

endo or exo5-hexyl-4-phenyl-3a-(1-phenylvinyl)-1,2,3,3a,6,6a-hexahydropentalene-1-carboxamide(38 endo, 38 exo)

To aflame dried round bottom flask was added potassium hydroxide (10.0eq) which was suspended in THF (0.1 M). The required nitrile isomer (32endo or 32 exo) (1.0 eq) was then added in THF dropwise and theresulting solution was heated to 65° C. for 48 hours. The solution wasthen washed with water (3×) and EtOAc (3×). The combined organic layerswere then washed with water and brine. The resulting solution was driedwith Na2SO4 and evaporated to dryness. The oil was then purified viacolumn chromatography in EtOAc/hexanes.

Endo ¹H NMR (400 MHz, Chloroform-d) δ 7.35-7.18 (m, 10H), 5.12 (m, 1H),5.02 (in, 1H), 3.60 (dt, J=18.3, 6.6 Hz, 1H), 2.77-2.66 (m, 1H),2.40-2.16 (n, 1H), 2.11-1.99 (m, 2H), 1.93-1.75 (m, 1H), 1.66-1.56 (m,2H), 1.47-1.23 (m, 4H), 0.97-0.76 (m, 3H). LCMS (90% MeCN/H2O): 2.15min, m/z 414.2.

Exo ¹H NMR (400 MHz, Chloroform-d) δ 7.35-7.18 (m, 10H), 5.12 (m, 1H),5.02 (m, 1H), 3.43 (bs, 1H), 2.40-2.16 (m, 1H), 2.11-1.99 (m, 4H),1.93-1.75 (m, 6H), 1.47-1.23 (m, 10H), 0.84 (td, J=7.2, 1.1 Hz, 3H).LCMS (90% MeCN/H2O): 2.34 min, m/z 414.7.

(endo orexo)-5-hexyl-4-phenyl-3a-(1-phenylethyl)-1,2,3,3a,6,6a-hexahydropentalen-1-ol(39 endo, 39 exo)

A flame dried round bottom flask charged with Pd/C (10 wt %) and wasplaced under argon. The solid was suspended in EtOAc (0.1 M) and theappropriate diastereomer of RJW100 (1.0 eq) was added dropwise in EtOAc(0.1 M). The resulting solution was backfilled with hydrogen and stirredfor 18 hours. The mixture was then filtered through a plug of silica andevaporated to dryness. The resulting oil was purified via columnchromatography in 5-10% EtOAc/hexanes.

Endo ¹H NMR (600 MHz, Chloroform-d) δ 7.18-6.99 (p, J=2.2 Hz, 6H),6.98-6.93 (m, 2H), 6.58-6.54 (m, 2H), 4.49 (dt, J=11.6, 6.6 Hz, 1H),2.68 (d, J=6.2 Hz, 1H), 2.60-2.55 (m, 1H), 2.53-2.46 (m, 1H), 2.18-2.08(m, 2H), 2.04 (dt, J=11.8, 7.5 Hz, 1H), 1.68-1.57 (m, 2H), 1.37 (dd,J=14.0, 9.6 Hz, 2H), 1.18 (d, J=6.3 Hz, 3H), 1.15-0.96 (m, 6H), 0.77 (t,J=7.3 Hz, 3H). Endo ¹³C NMR (600 MHz, Chloroform-d) δ 145.77, 141.05,128.35, 127.33, 127.28, 125.50, 125.36, 77.22, 75.72, 64.65, 60.06,53.58, 44.25, 44.01, 33.86, 32.73, 32.07, 31.76, 30.37, 29.71, 29.44,28.50, 22.52, 20.11, 14.03.

Exo ¹H NMR (600 MHz, Chloroform-d) δ 7.08-7.00 (m, 3H), 6.95-6.88 (m,3H), 6.68 (d, J=7.3 Hz, 2H), 6.58 (d, J=7.3 Hz, 2H), 4.20 (dt, J=5.0,2.2 Hz, 1H), 2.77 (d, J=6.5 Hz, 1H), 2.53 (d, J=10.5 Hz, 1H), 2.24-1.98(m, 5H), 1.80 (qd, J=8.0, 7.5, 3.4 Hz, 1H), 1.64 (ddd, J=13.6, 9.1, 4.5Hz, 2H), 1.48 (dd, J=13.7, 7.1 Hz, 5H), 1.32 (d, J=6.7 Hz, 3H),1.16-0.94 (m, 9H), 0.85 (t, J=7.1 Hz, 3H). Exo ¹³C NMR (400 MHz,Chloroform-d) δ 146.99, 140.97, 130.62, 128.24, 127.14, 127.03, 125.20,124.75, 83.37, 77.19, 64.16, 62.14, 57.43, 45.46, 42.60, 38.19, 36.77,36.36, 31.72, 31.29, 29.93, 29.69, 29.39, 28.42, 22.49, 21.08, 14.01.

((((endo or exo)-5-hexyl-4-phenyl-3a-(1-phenylvinyl)-1,2,3,3a,6,6a-hexahydropentalen-1-yl)oxy)carbonyl)sulfamicacid (40 endo, 40 exo)

In a flame dried round bottom flask was added the appropriate isomer (29endo, or 29 exo) in ACN (0.1 M). Chlorosulfonyl isocyanate (2.0 eq) wasadded after cooling the solution to −15° C. After stirring around 10minutes, water (0.04 M) was added and heated at 60° C. for around 18hours. After cooling to room temperature, the resulting solution wasquenched with water and adjusted to pH 5. The resulting solution wasextracted with EtOAc (3×). The combined organic layers were then driedwith Na2SO4 and evaporated to dryness. The resulting oil was purifiedvia column chromatography in 50% EtOAc/hex.

Endo ¹H NMR (400 MHz, Chloroform-d) δ 7.32-7.17 (m, 11H), 5.05 (d, J=1.4Hz, 1H), 4.96-4.87 (m, 2H), 4.62 (s, 1H), 2.66 (td, J=8.9, 1.9 Hz, 1H),2.33 (dd, J=16.5, 1.4 Hz, 1H), 2.10-1.96 (m, 5H), 1.93-1.86 (m, 1H),1.70-1.60 (m, 3H), 1.31-1.18 (m, 9H), 0.89-0.84 (m, 3H).

Exo ¹H NMR (400 MHz, Chloroform-d) δ 7.32-7.18 (m, 10H), 5.04 (d, J=1.5Hz, 1H), 4.98 (d, J=1.5 Hz, 1H), 4.75-4.70 (m, 1H), 2.37 (td, J=8.9, 1.1Hz, 1H), 2.34-2.27 (m, 1H), 2.18-2.15 (m, 1H), 2.04-1.91 (m, 6H),1.78-1.65 (m, 3H), 1.36-1.15 (m, 9H), 0.84 (t, J=7.0 Hz, 3H).

(endo orexo)-5-hexyl-4-phenyl-3a-(1-phenylvinyl)-1,2,3,3a,6,6a-hexahydropentalen-1-ylsulfamoylcarbamate (41 endo, 41 exo)

In a flame dried round bottom flask was added 6 in ACN (0.1 M) andchlorosulfonyl isocyanate (2.0 eq) was subsequently added after coolingthe solution to −15° C. After stirring around 10 minutes, ammoniumhydroxide (0.04 M) was added and heated at 60° C. for around 18 hours.After cooling to room temperature, the resulting solution was quenchedwith a saturated solution of bicarbonate to pH 9. The resulting solutionwas extracted with EtOAc (3×). The combined organic layers were thendried with Na2SO4 and evaporated to dryness. The resulting oil waspurified via column chromatography in 50% EtOAc/hex with 1%triethylamine.

Exo ¹H NMR (500 MHz, Chloroform-d) δ 7.36-7.28 (m, 8H), 7.22-7.18 (m,2H), 5.16 (s, 2H), 5.12 (d, J=1.3 Hz, 1H), 5.00 (d, J=1.3 Hz, 1H), 4.91(d, J=3.6 Hz, 1H), 2.48-2.41 (m, 2H), 2.21-2.13 (m, 1H), 2.04 (ddd,J=8.6, 3.9, 2.0 Hz, 2H), 1.98-1.75 (m, 5H), 1.39-1.18 (m, 9H), 0.87 (t,J=7.1 Hz, 3H).

Endo ¹H NMR (500 MHz, Chloroform-d) δ 7.35-7.19 (m, 10H), 5.22 (s, 2H),5.12-5.04 (m, 2H), 4.97 (d, J=1.3 Hz, 1H), 2.71 (dd, J=8.8, 1.9 Hz, 1H),2.30 (dd, J=17.3, 1.9 Hz, 1H), 2.12-2.04 (m, 2H), 1.97 (q, J=6.3 Hz,1H), 1.81-1.69 (m, 3H), 1.40-1.20 (m, 9H), 0.94-0.83 (t, 3H).

5-hexyl-4-phenyl-3a-(1-phenylvinyl)-3,3a,6,6a-tetrahydropentalen-1(2H)-oneoxime (42)

A flame dried round bottom flask was charged with hydroxylaminehydrochloride (1.2 eq) and sodium acetate (1.3 eq) before beingbackfilled with argon. The solids were dissolved in an EtOH/watermixture (4:1, 0.1 M). 27 (1.0 eq) in EtOH was added dropwise and heatedat 80° C. for 6 hours. The solution was then washed with water (3×) andEtOAc (3×). The combined organic layers were then washed with water andbrine. The resulting solution was dried with Na2SO4 and evaporated todryness. The oil was then purified via column chromatography in 5%EtOAc/hexanes.

1-ethynyl-5-hexyl-4-phenyl-3a-(1-phenylvinyl)-1,2,3,3a,6,6a-hexahydropentalen-1-ol(43)

A solution of 27 (1.0 eq) in dry THE was cooled to −78° C. in a flamedried round bottom flask under nitrogen. Ethynyl magnesium bromide (0.5M, 1.5 eq) was then added dropwise. The resulting solution was stirredfor 20 hours to room temperature. Ammonium chloride was added to thesolution and extracted with EtOAc (3×). The combined organic layers weredried with Na2SO4 and evaporated to dryness. The resulting oil waspurified in 0-10% EtOAc/hex.

Endo ¹H NMR (400 MHz, Chloroform-d) δ 7.34-7.14 (m, 10H), 5.06 (s, 1H),5.00 (d, J=1.4 Hz, 1H), 4.11 (qd, J=7.1, 1.1 Hz, 1H), 2.73-2.58 (m, 2H),2.44 (d, J=0.8 Hz, 1H), 2.12-1.81 (m, 5H), 1.74 (dt, J=12.2, 5.4 Hz,1H), 1.36 (q, J=7.4 Hz, 2H), 1.32-1.12 (m, 9H), 0.84 (t, J=6.8 Hz, 3H).¹³C NMR (101 MHz, Chloroform-d) δ 154.45, 143.66, 142.53, 139.19,136.87, 129.76, 127.90, 127.69, 127.67, 126.73, 126.65, 114.90, 88.50,75.17, 71.45, 68.71, 57.12, 39.84, 34.46, 32.40, 31.63, 29.78, 29.42,27.86, 22.58, 21.06, 14.19, 14.08.

(endo or exo)(5-hexyl-4-phenyl-3a-(1-phenylvinyl)-1,2,3,3a,6,6a-hexahydropentalen-1-10yl)methanamine (48)

To an oven-dried reaction tube under nitrogen was added lithium aluminumhydride. After being suspended in ether, the appropriate diastereomer of32 (1.0 equiv.) in ether was added dropwise. The reaction mixture wasallowed to stir for 18 h before being quenched with water and 1M NaOHbefore being filtered through celite and evaporated in vacuo. Theresulting crude oil was purified on basified silica in 10% MeOH/DCM(endo or exo)5-hexyl-4-phenyl-3a-(1-phenylvinyl)-1,2,3,3a,6,6a-hexahydropentalen-1-yldihydrogen phosphate (49)

In a reaction vial under nitrogen, RJW100 (1.0 equiv.) was dissolved inacetonitrile. To the reaction vial was added trimethylamine (4.0 equiv.)and phosphoryl chloride (2.0 equiv.) The reaction was stirred at roomtemperature under nitrogen for 4 h before being quenched with water(excess) and concentrated. The resulting crude residue was purified onbasified silica in 10% MeOH/DCM.

(endo orexo)-5-hexyl-4-phenyl-3a-(1-phenylvinyl)-1,2,3,3a,6,6a-hexahydropentalen-1-ylcarbamoylsulfamate (50)

In a reaction tube under nitrogen was added NaH (1.5 equiv.) in THF. Thesolution was cooled to 0 C before the addition of a solution of therequired sulfamate (28 endo, 28 exo) (1.0 equiv.) in THF. A solution ofcarbonyldiimidazole (2.0 equiv.) in THE was added and the reactionmixture was allowed to warm to room temperature. After stirring for 1 h,ammonia (7N in MeOH) was added dropwise and stirred for ˜3 h. Afterallowing to stir, the reaction was diluted with EtOAc and washed withbrine (ensuring the aqueous layer was basic). The organic layer wasdried with MgSO4, filtered, and concentrated in vacuo. The crude oil waspurified on silica in 50% EtOAc to 10% MeOH/DCM.

Endo: ¹H NMR (400 MHz, Chloroform-d) δ 7.35 (s, 2H), 7.32-7.25 (m, 4H),7.24-7.21 (m, 2H), 7.16 (ddd, J=7.2, 3.6, 1.8 Hz, 2H), 5.08 (d, J=1.2Hz, 1H), 5.06-4.97 (m, 1H), 4.93-4.89 (m, 1H), 4.89-4.80 (m, 1H), 3.74(s, 2H), 2.67 (td, J=8.8, 2.3 Hz, 1H), 2.62-2.48 (m, 1H), 2.12 (ddd,J=17.8, 9.1, 3.9 Hz, 1H), 2.06-2.00 (m, 1H), 1.76 (ddd, J=8.5, 6.4, 3.4Hz, 1H),1.67 (td, J=12.6, 5.7 Hz, 1H), 1.35 (d, J=6.4 Hz, 4H), 1.28-1.16(m, 7H), 0.83 (t, J=7.0 Hz, 3H). LRMS [ESI-APCI] calc'd for C28H33[M-CH3N2O4S]+369.3, found 369.2

Exo: ¹H NMR (400 MHz, Chloroform-d) δ 7.28-7.25 (m, 2H), 7.24-7.22 (m,1H), 7.20-7.12 (m, 5H), 5.39 (d, J=48.0 Hz, 1H), 4.98 (s, 1H), 4.93 (s,1H), 4.69 (s, 1H), 3.54 (s, 3H), 2.64-2.55 (m, 1H), 2.33-2.18 (m, 1H),1.74-1.57 (m, 2H), 1.31-1.10 (m, 14H), 0.81 (t, J=7.1 Hz, 3H). Exo LRMS[ESI-APCI] calc'd for C28H33 [M-CH3N2O4S]+ 369.3, found 369.2

5′-hexyl-4′-phenyl-3a′-(1-phenylvinyl)-3′,3a′,6′,6a′-tetrahydro-2′H-spiro[oxirane-52,1′-pentalene]

Trimethylsilyliodide (3.0 equiv.) was dissolved in a reaction vial opento air. Potassium Hydroxide (12.0 equiv.) was added and stirred untildissolved, then water (˜20 equiv.) was added. 27 in acetonitrile wasadded to the reaction mixture before the reaction was heated to 60° C.for 16 h. The reaction mixture was then cooled to room temperature andfiltered through celite. The filtrate was then poured onto water andextracted three times with EtOAc. The organic layers were concentratedin vacuo and the crude oil was purified on silica in 5-10% EtOAc/Hex.

3-(aminomethyl)-5-hexyl-6-phenyl-6a-(1-phenylvinyl)-1,2,3,3a,4,6a-hexahydropentalen-2-ol(51)

In a reaction tube under nitrogen, the prepared expoxide5′-hexyl-4′-phenyl-3a′-(1-phenylvinyl)-3′,3a′,6′,6a′-tetrahydro-2′H-spiro[oxirane-52,1′-pentalene] (1.0 equiv.) was dissolved in acetonitrile. Ammonia inmethanol (7N, excess) was added. The reaction tube was capped and heatedto 60 C for 18 h. The reaction mixture was then cooled to roomtemperature and concentrated in vacuo. The resulting crude oil waspurified on silica in 10% MeOH/DCM.((5-hexyl-4-phenyl-3a-(1-phenylvinyl)-3,3a,6,6a-tetrahydropentalen-1-yl)oxy)triethylsilaneUnder nitrogen, freshly prepared lithium diisopropylamide (1.1 equiv.)was added to THE and the solution was cooled to −78 C and 27 (1.0equiv.) in TH was added. The reaction mixture was stirred at −78 C for20 minutes before the addition of triethylsilyl chloride (2.1 equiv.).The reaction mixture was stirred for 1.5 h and allowed to warm to roomtemperature before being quenched with ammonium chloride. The crudemixture was then extracted with ethyl acetate. The organic layers werewashed with brine, dried with magnesium sulfate, and concentrated invacuo. The resulting crude oil was purified on silica in 5% EtOAc/Hex.

(exo or endo)5-hexyl-2-hydroxy-4-phenyl-3a-(1-phenylvinyl)-3,3a,6,6a-tetrahydropentalen-1(2H)-one

Sodium bicarbonate (5.0 equiv.) and meta-chloroperoxybenzoic acid (1.2equiv.) were added in a reaction tube and backfilled with nitrogen.Hexanes was added and the resulting suspension was cooled to −15° C.((5-hexyl-4-phenyl-3a-(1-phenylvinyl)-3,3a,6,6a-tetrahydropentalen-1-yl)oxy)triethylsilane(1.0 equiv.) in hexane was added to the reaction tube and the resultingmixture was allowed to stir for 1 h before being poured onto water andextracted with ethyl acetate. The organic layers were washed with brine,dried with magnesium sulfate and concentrated in vacuo. The resultingcrude oil was purified in 10% EtOAc/Hex.

5-hexyl-4-phenyl-3a-(1-phenylvinyl)-1,2,3,3a,6,6a-hexahydropentalene-1,2-diol(52)

(exo or endo)5-hexyl-2-hydroxy-4-phenyl-3a-(1-phenylvinyl)-3,3a,6,6a-tetrahydropentalen-1(2H)-one(1.0 equiv.) was dissolved in methanol and sodium borohydride (excess)was added. The reaction mixture was stirred until effervescence ceased.The crude reaction mixture was poured onto water and extracted withEtOAc, dried with magnesium sulfate and concentrated in vacuo. Theresulting crude oil was purified on silica in 50% EtOAc/Hex.

Synthesis of Additional Derivatives

The synthetic procedure for preparing compounds 44, 45a, 45b, 46a, 46b,and 45c is provided for in FIG. 8.

10-(6-oxo-3-phenyl-3a-(1-phenylvinyl)-1,3a,4,5,6,6a-hexahydropentalen-2-yl)decanoicacid (44)

A solution of 1f (1.0 equiv.) in acetonitrile was treated withN-methylmorpholine oxide (1.5 equiv.) and allowed to stir to homogeneitybefore the addition of tetrapropylammonium perruthenate (0.1 equiv.) Thesolution was stirred at room temperature until completion as determinedby TLC (˜10 min). The solution was concentrated in vacuo and subjecteddirectly to silica gel chromatography in 10% EtOAc/Hex to afford thetitle compound as a clear, colorless oil.

10-(endo)-6-hydroxy-3-phenyl-3a-(1-phenylvinyl)-1,3a,4,5,6,6a-hexahydropentalen-2-yl)decanoicacid (45a)

In a scintillation vial, 44 was dissolved in MeOH. Sodium borohydride(1.1 equiv.) was added, and the reaction was allowed to stir tocompletion (30 min). The mixture was then concentrated and passedthrough a short plug of silica (eluted with EtOAc). The reaction isselective for the endo diastereomer shown in 45a.

10-(endo)-3-phenyl-3a-(1-phenylvinyl)-6-(sulfamoyloxy)-1,3a,4,5,6,6a-hexahydropentalen-2-yl)decanoicacid (45b)

In a scintillation vial, 45a was dissolved in DMA. Sulfamoyl chloride(1.1 equiv.) was added and allowed to stir for 1 hour before theaddition of triethylamine (1.1 equiv.). The reaction mixture was thenallowed to stir for an additional 10 minutes. The resulting solution wasconcentrated and purified on silica in 20-100% EtOAc/Hex (1% aceticacid).

10-(exo)-3-phenyl-3a-(1-phenylvinyl)-6-(sulfamoyloxy)-1,3a,4,5,6,6a-hexahydropentalen-2-yl)decanoicacid (45c)

In a scintillation vial, 1f was dissolved in DMA. Sulfamoyl chloride(1.1 equiv.) was added and allowed to stir for 1 hour before theaddition of triethylamine (1.1 equiv.) The reaction mixture was thenallowed to stir for an additional 10 minutes. The resulting solution wasconcentrated and purified on silica in 20-100% EtOAc/Hex (1% aceticacid).

¹H NMR (500 MHz, Chloroform-d) δ 7.38-7.13 (m, 10H), 5.08 (d, J=1.5 Hz,1H), 4.99 (d, J=1.5 Hz, 1H), 4.83 (s, 2H), 4.72 (d, J=4.2 Hz, 1H), 2.66(d, J=9.3 Hz, 1H), 2.35 (t, J=7.4 Hz, 3H), 2.12 (d, J=17.6 Hz, 1H),2.09-1.93 (m, 7H), 1.87-1.71 (m, 2H), 1.70-1.58 (m, 3H), 1.39-1.10 (m,10H). ¹³C NMR (126 MHz, cdcl3) δ 176.48, 153.48, 143.61, 141.35, 138.91,136.81, 129.53, 127.81, 127.78, 127.72, 126.88, 115.57, 93.70, 69.29,52.81, 40.13, 33.32, 32.08, 32.06, 29.43, 29.24, 28.87, 28.83, 28.79,28.75, 27.51, 24.53. FT-IR (neat): 3475, 3275, 3080, 3053, 3019, 2924,2853, 1707, 1598, 1572, 1491, 1457, 1440, 1357, 1260, 1181, 1092, 1075,1027, 927, 904, 800, 763, 701 cm-1.

10-(endo)-6-amino-3-phenyl-3a-(1-phenylvinyl)-1,3a,4,5,6,6a-hexahydropentalen-2-yl)decanoicacid (46a)

In a reaction vial equipped with stir bar, 44 was dissolved in methanol.Titanium tetraisopropoxide (TiO(iPr)4), (2.0 equiv.) was added portionwise. Ammonia in Methanol (7N) was then added to the reaction mixtureand allowed to stir at room temperature for 6 h. Sodium borohydride (2.0equiv.) was added slowly, and the reaction mixture was stirred untileffervescence ceased. The resulting mixture was concentrated andpurified on a plug of silica in 65/35/5 DCM/MeOH/NH4OH.

10-(endo)-3-phenyl-3a-(1-phenylvinyl)-6-(sulfamoylamino)-1,3a,4,5,6,6a-hexahydropentalen-2-yl)decanoicacid (46b)

To a solution of the required amine (44) (1.0 equiv.) and triethylamine(2.0 equiv.) in DCM under nitrogen was added a 0.5M solution of2-oxo-1,3-oxazolidine-3-sulfonyl chloride in DCM (1.0 equiv.) (preparedaccording to the procedure of Borghese et al.). The resulting solutionwas stirred at room temperature for three hours then concentrated invacuo to a solid. To the resulting crude solid was added ammonia (0.5Min dioxane, 1.5 equiv.) and triethylamine (3.0 equiv.). The solution wasthen heated in a sealed tube to 85° C. for 4 h. After cooling to roomtemperature, the reaction was diluted with 3:3:94 MeOH:Et3N:EtOAc thenpassed through a pad of silica. The eluent was concentrated in vacuo andpurified by silica gel chromatography in 50-100% EtOAc/hexanes (1%Acetic Acid) to afford the title compound as a colorless oil.

(exo or endo)8-(6-hydroxy-3-phenyl-3a-(1-phenylvinyl)-1,3a,4,5,6,6a-hexahydropentalen-2-yl)octylacetate (47)

In an oven-dried vial under nitrogen, the required diol (1.0 equiv.) wasdissolved in DCM and cooled to 0° C. Triethylamine (2.0 equiv.) wasadded to the reaction mixture before acetyl chloride (1.0 equiv.). Themixture was allowed to warm to room temperature and stir for 30 minbefore being concentrated in vacuo. The crude oil was purified on aglass-backed preparative alumina plate in 30% EtOAc/Hex.

(endo or exo) diethyl(8-(6-hydroxy-3-phenyl-3a-(1-phenylvinyl)-1,3a,4,5,6,6a-hexahydropentalen-2-yl)octyl)phosphate (53)

In an oven-dried vial, the required alcohol isomer (1.0 equiv.) wasdissolved in DCM. Triethylamine (2.0 equiv.) was added and cooled to 0°C. before diethyl chlorophosphate (1.0 equiv.) was added. The reactionmixture was stirred at room temperature for 18 h before beingconcentrated in vacuo and purified on a glass-backed preparative aluminaplate in 30% EtOAc/Hex.

Biological Evaluation of Novel LRH-1 Agonists and Antagonists.

Protein expression and purification-LRH-1 LBD (residues 299-541) in thepMSC7 vector was expressed in BL21(DE3) pLysS E. coli by induction withIPTG (1 mM) for 4 hr at 30° C. Protein was purified by nickel affinitychromatography. Protein used for Thermofluor experiments was incubatedwith DLPC (five-fold molar excess) for four hours at room temperature,and then repurified by size-exclusion into an assay buffer of 20 mMTris-HCl, pH 7.5, 150 mM NaCl, and 5% glycerol. Protein used forcrystallization was incubated with TEV protease to cleave the His tag.The cleaved protein was then separated from the His tag and TEV by asecond round of nickel affinity chromatography. To make protein-ligandcomplexes, protein was incubated with ligands overnight (10-fold molarexcess) and repurified by size-exclusion, using a final buffer of 100 mMammonium acetate, pH 7.4, 150 mM sodium chloride, 1 mM DTT, 1 mM EDTA,and 2 mM CHAPS.

Thermofluor assays-Purified LRH-1 LBD-His protein (0.2 mg/ml) wasincubated overnight with 50 M of each compound at 4° C. The final DMSOconcentration in the reactions was 1%. SYPRO orange dye (Invitrogen) wasthen added at a 1:1000 dilution. Reactions were heated at a rate of 0.5°C. per minute, using a StepOne Plus Real Time PCR System (ThermoFisher).Fluorescence was recorded at every degree using the ROX filter (602 nm).Data were analyzed by first subtracting baseline fluorescence(ligands+SYPRO with no protein) and then fitting the curves using theBolzman equation (GraphPad Prism, v6) to determine the Tm.

Luciferase reporter assays-HeLa cells were seeded at a density of 10,000cells per well in white-walled, clear-bottomed 96-well culture plates.The next day, cells were transfected with LRH-1 and reporters, usingFugene HD (Roche) at a ratio of 5:2 Fugene (μl): DNA (g). Thetransfected plasmids included full-length LRH-1 in a pCI vector (5ng/well), and a SHP-luc reporter, encoding the LRH-1 response elementand surrounding sequence from the SHP promoter cloned upstream offirefly luciferase in the pGL3 basic vector (50 ng/well). Cells werealso co-transfected with a constitutive Renilla luciferase reporter(utilizing the CMV promoter), which was used for normalization offirefly signal (1 ng/well). Control cells received pCI empty vector at 5ng/well in place of LRH-1-pCI. Following an overnight transfection,cells were treated with agonists for 24 hours at the concentrationsindicated in the figure legends. Agonists were dissolved in DMSO andthen diluted into media, with a final concentration of 0.3% DMSO in allwells. Luciferase signal was quantified using the DualGlo kit (Promega).Experiments were conducted at least two times in triplicate.

Compounds were synthesized and evaluated in biological assays.Modifications to the scaffold were introduced at the RJW100 hydroxylsite, at the styrene ring, the other phenyl ring, and at other sites onthe core of the molecule. Notably, the substitution of the hydroxyl witha sulfamino moiety (50-endo/exo) greatly stabilized the LRH-1-ligandcomplex with comparable or slightly higher LRH-1 activation. The crystalstructure of LRH-1 bound to 50-endo was determined, which demonstratesthat the ligand displaces the water molecule normally coordinated byLRH-1 residue T352 to directly interact with this threonine. Thecompound 40-endo also greatly stabilized the protein-ligand complex inThermofluor assays and demonstrated increased activation of LRH-1compared to RJW100. Compound 48-endo showed dose-dependent inhibition ofLRH-1 activity.

A second strategy for improvement of activity involved modifications tothe tail of the molecule. The effects of tail length and addition ofvarious polar groups to the end of the tail were explored. Tail lengthsof 8-10 carbons in length conferred the strongest activation of LRH-1.

Methods

Protein expression and purification—LRH-1 LBD (residues 299-541) in thepMSC7 vector was expressed in BL21(DE3) pLysS E. coli by induction withIPTG (1 mM) for 4 hr at 30° C. Protein was purified by nickel affinitychromatography. Protein used for thermofluor experiments was incubatedwith DLPC (five-fold molar excess) for four hours at room temperature,and then repurified by size-exclusion into an assay buffer of 20 mMTris-HCl, pH 7.5, 150 mM NaCl, and 5% glycerol. Protein used forcrystallization was incubated with TEV protease to cleave the His tag.The cleaved protein was then separated from the His tag and TEV by asecond round of nickel affinity chromatography. To make protein-ligandcomplexes, protein was incubated with ligands overnight (10-fold molarexcess) and repurified by size-exclusion, using a final buffer of 100 mMammonium acetate, pH 7.4, 150 mM sodium chloride, 1 mM DTT, 1 mM EDTA,and 2 mM CHAPS.

Thermofluor assays—Purified LRH-1 LBD-His protein (0.2 mg/ml) wasincubated overnight with 50 M of each compound at 4° C. The final DMSOconcentration in the reactions was 1%. SYPRO orange dye (Invitrogen) wasthen added at a 1:1000 dilution. Reactions were heated at a rate of 0.5°C. per minute, using a StepOne Plus Real Time PCR System (ThermoFisher).Fluorescence was recorded at every degree using the ROX filter (602 nm).Data were analyzed by first subtracting baseline fluorescence(ligands+SYPRO with no protein) and then fitting the curves using theBolzman equation (GraphPad Prism, v6) to determine the Tm.

Luciferase reporter assays—HeLa cells were seeded at a density of 10,000cells per well in white-walled, clear-bottomed 96-well culture plates.The next day, cells were transfected with LRH-1 and reporters, usingFugene HD (Roche) at a ratio of 5:2 Fugene (μl): DNA (μg). Thetransfected plasmids included full-length LRH-1 in a pCI vector (5ng/well), and a SHP-luc reporter, encoding the LRH-1 response elementand surrounding sequence from the SHP promoter cloned upstream offirefly luciferase in the pGL3 basic vector (50 ng/well). Cells werealso co-transfected with a constitutive Renilla luciferase reporter(utilizing the CMV promoter), which was used for normalization offirefly signal (1 ng/well). Control cells received pCI empty vector at 5ng/well in place of LRH-1-pCI. Following an overnight transfection,cells were treated with agonists for 24 hours at concentrations of 0.03,0.3, 1, 3, 10, and 30 μM of each compound. Agonists were dissolved inDMSO and then diluted into media, with a final concentration of 0.3%DMSO in all wells. Luciferase signal was quantified using the DualGlokit (Promega).

Crystallization—Protein-ligand complexes were incubated with a peptidederived from human Tif2 NR Box 3 or SMRT at four-fold molar excess fortwo hours at room temperature and then concentrated to 6.5 mg/ml.Crystallization conditions were identified using our Phoenix robot toscreen in a high throughput manner or were based on the conditions usedto produce RJW100-LRH-1 crystals.

Summary of Biological Data from Compound Screens

EC₅₀ and Relative Activity values were calculated from luciferasereporter assays. Relative activity refers to fold increase in activityover baseline (DMSO-treated cells) for each compound relative to thefold increase induced by RJW100. A value of “0” indicates no activityover baseline, values between 0 and 1 indicate less active compounds,and values over 1 indicate compounds that are more active than RJW100 inthese assays. ΔTm values were determined in thermofluor assays and referto the difference in melting temperature of the LRH-1 ligand complexrelative to DLPC-bound LRH-1 (PC 12:0/12:0, aphospholipid LRH-1agonist). The Tm of RJW100 relative to DLPC is ˜3° C.

Diols, Phosphorylcholines, Carboxylic Acids

Compound Relative Number EC₅₀ (μM) Efficacy ΔTm (mean +/− SEM) (° C.) 5e 1.4 +/− 0.5 0.83 3.1 +/− 0.9  6e cnc 1.57 2.4 +/− 0.3  7e 1.0 +/−0.8 1.74 3.3 +/− 0.8  8e 0.2 +/− 0.2 1.26 3.7 +/− 0.8  9e 0.7 +/− 0.91.52 4.0 +/− 0.5 10e 1 +/− 2 1.30 4.1 +/− 0.6 11e 2 +/− 5 2.83 ns 12e 2+/− 2 0.87 4.7 +/− 0.9  5g cnc 0.33 ns  6g cnc 0.00 ns  7g cnc 0.00 ns 8g cnc 0.81 2.9 +/− 0.4  9g >30 1.12 5.6 +/− 0.6 10g 2 +/− 3 2.74 ns11g 5 +/− 2 2.48 −2.4 +/− 0.5   12g 5.1 +/− 0.5 0.93 −1.7 +/− 0.9    5fcnc 1.43 2.3 +/− 0.2  6f cnc 0.60 1.3 +/− 0.4  7f 9 +/− 3 2.22 5.5 +/−0.3  8f 4 +/− 3 1.17 3.7 +/− 0.2  9f 4 +/− 2 2.48 4.4 +/− 0.5 10f 1.8+/− 0.7 3.39 3 +/− 1 11f 0.4 +/− 0.4 2.39 ns 12f 0.3 +/− 0.3 1.74 2.5+/− 0.3

Other Tail Modifications and Hybrids

Compound Relative Number EC₅₀ (μM) Efficacy ΔTm (mean +/− SEM) (° C.)13b 12 +/− 6  0.96 nd 45b 0.07 +/− 0.3  0.66 8.4 +/− 0.2 45c 0.2 +/− 0.11.85 ns 46b >30 0.51 8.5 +/− 0.2 47-endo nd 1.48 ns 47-exo 0.7 +/− 0.73.70 1.5 +/− 0.2 51 cnc 0.65 nd 52 0.01 +/− 0.03 −0.25 nd 53-endo nd1.04 ns 53-exo nd 0.37 ns 54 16 +/− 11 1.05 nd 14d 17 +/− 7  3.35 nd

Modifications at RJW100 Internal Styrene

Compound Relative Number EC₅₀ (μM) Efficacy ΔTm (mean +/− SEM) (° C.)15f cnc 0.85 1.4 +/− 0.1 15g cnc 0.70 ns 16f >30 0.9 ns 16g 24 +/− 473.8 ns 17f 3 +/− 4 0.6 −1.9 +/− 0.5   17g >30 2.7 ns 18f >30 2.8 ns 18gcnc 0.99 1.5 +/− 0.2

Modifications at RJW100 External Styrene

Compound Relative Number EC₅₀ (μM) Efficacy ΔTm (mean +/− SEM) (° C.) 204+/−4 0.85 1.4 +/− 0.7 21 0.9 +/− 0.5 0.87 1.3 +/− 0.2 22 cnc 0.42 ns 24cnc −0.14 −3.1 +/− 0.8   23a cnc 0.06 nd 23b 5 +/− 9 0.86 ns 25-endo cnc0.42 ns 25-exo 0.8 +/− 0.7 0.31 1.0 +/− 0.4 26-endo 0.4 +/− 1   0.11 1.9+/− 0.1 26-exo 9 +/− 4 1.41 1.4 +/− 0.1

Modifications at RJW100 Hydroxyl Site

Compound Relative Number EC₅₀ (μM) Efficacy ΔTm (mean +/− SEM) (° C.) 27nd −0.42 ns 28-endo 0.9 +/− 0.6 0.96 8 +/− 2 28-exo 0.8 +/− 0.6 0.62 5.1+/− 0.2 29-endo cnc 0 2.8 +/− 0.3 29-exo cnc 0.14 1.54 +/− 0.04 30-endo0.5 +/− 0.2 0.72 3 +/− 1 30-exo 1 +/− 1 0.14 1.1 +/− 0.4 31-endo 5 +/− 50.70 ns 31-exo 2 +/− 3 0.45 ns 32-endo nd 0 ns 32-exo nd 0 ns 33-endo nd0.56 ns 33-exo nd 0.56 −3.0 +/− 0.8   34-endo 1 +/− 1 1.2 3.6 +/− 0.434-exo cnc cnc 0.6 +/− 0.3 35-endo 13 +/− 8  2.6 0.4 +/− 0.4 35-exo 17−0.1 −1.1 +/− 0.6   36-endo 0.5 +/− 0.7 0.30 1.1 +/− 0.3 36-exo 2 +/− 30.41 −1.4 +/− 0.4   37-endo 0.015 +/− 0.008 1.3 9.2 +/− 0.5 37-exo 0.04+/− 0.05 0.39 4.1 +/− 0.3 39-endo nd 0.13 0.4 +/− 0.9 39-exo nd 0.06 0.1+/− 1   40-endo >30 −0.2 1.95 +/− 0.15 40-exo >30 0 nd 48-endo cnc 0 0Cnc = could not calculate EC50 Nd = not done Ns = Tm changes were lessthan ° C.

1. A method of treating or preventing cardiovascular disease comprisingadministering to a subject in need thereof an effective amount of acompound having the following formula:

prodrugs or salts thereof wherein, R¹ is alkyl, halogen, nitro, cyano,hydroxy, amino, mercapto, formyl, carboxy, carbamoyl, alkoxy,hydroxyalkyl, alkylthio, thioalkyl, alkylamino, aminoalkyl,(alkyl)₂amino, alkanoyl, alkoxycarbonyl, alkylsulfinyl, alkylsulfonyl,arylsulfonyl, carbocyclyl, benozyl, benzyl, aryl, or heterocyclyl,wherein R¹ is optionally substituted with one or more, the same ordifferent, R¹⁰; R² is hydrogen, alkyl, halogen, nitro, cyano, hydroxy,amino, mercapto, formyl, carboxy, carbamoyl, alkoxy, hydroxyalkyl,alkylthio, thioalkyl, alkylamino, aminoalkyl, (alkyl)₂amino, alkanoyl,alkoxycarbonyl, alkylsulfinyl, alkylsulfonyl, arylsulfonyl, carbocyclyl,benozyl, benzyl, aryl, or heterocyclyl, wherein R² is optionallysubstituted with one or more, the same or different, R¹⁰; R³ ishydrogen, alkyl, halogen, nitro, cyano, hydroxy, amino, mercapto,formyl, carboxy, carbamoyl, alkoxy, hydroxyalkyl, alkylthio, thioalkyl,alkylamino, aminoalkyl, (alkyl)₂amino, alkanoyl, alkoxycarbonyl,alkylsulfinyl, alkylsulfonyl, arylsulfonyl, carbocyclyl, benozyl,benzyl, aryl, or heterocyclyl, wherein R³ is optionally substituted withone or more, the same or different, R¹⁰; R⁴ is alkyl, halogen, nitro,cyano, hydroxy, amino, mercapto, formyl, carboxy, carbamoyl, alkoxy,hydroxyalkyl, alkylthio, thioalkyl, alkylamino, aminoalkyl,(alkyl)₂amino, alkanoyl, alkoxycarbonyl, alkylsulfinyl, alkylsulfonyl,arylsulfonyl, carbocyclyl, benozyl, benzyl, aryl, or heterocyclyl,wherein R⁴ is optionally substituted with one or more, the same ordifferent, R¹⁰; R⁵ is alkyl, halogen, nitro, cyano, hydroxy, amino,mercapto, formyl, carboxy, carbamoyl, alkoxy, hydroxyalkyl, alkylthio,thioalkyl, alkylamino, aminoalkyl, (alkyl)₂amino, alkanoyl,alkoxycarbonyl, alkylsulfinyl, alkylsulfonyl, arylsulfonyl, carbocyclyl,benozyl, benzyl, aryl, or heterocyclyl, wherein R⁵ is optionallysubstituted with one or more, the same or different, R¹⁰; R⁶ is lipid oralkyl, wherein R⁶ is optionally terminally substituted with a hydroxy,carboxy, or phosphate, wherein the hydroxy, carboxy, or phosphate areoptionally further substituted with R¹⁰; R⁷ is hydrogen, alkyl, halogen,nitro, cyano, hydroxy, amino, mercapto, formyl, carboxy, carbamoyl,alkoxy, hydroxyalkyl, alkylthio, thioalkyl, alkylamino, aminoalkyl,(alkyl)₂amino, alkanoyl, alkoxycarbonyl, alkylsulfinyl, alkylsulfonyl,arylsulfonyl, carbocyclyl, benozyl, benzyl, aryl, or heterocyclyl,wherein R⁷ is optionally substituted with one or more, the same ordifferent, R¹⁰; or R¹ and R⁷ together are an oxo or oxime, wherein theoxime is optionally substituted with one or more, the same or different,R¹⁰; R¹⁰ is alkyl, halogen, nitro, cyano, hydroxy, amino, mercapto,formyl, carboxy, carbamoyl, alkoxy, hydroxyalkyl, alkylthio, thioalkyl,alkylamino, aminoalkyl, (alkyl)₂amino, alkanoyl, alkoxycarbonyl,alkylsulfinyl, alkylsulfonyl, arylsulfonyl, aminosulfonyl, carbocyclyl,benozyl, benzyl, aryl, or heterocyclyl, wherein R¹⁰ is optionallysubstituted with one or more, the same or different, R¹¹; and R¹¹ ishalogen, nitro, cyano, hydroxy, trifluoromethoxy, trifluoromethyl,amino, formyl, carboxy, carbamoyl, mercapto, sulfamoyl, methyl, ethyl,methoxy, ethoxy, isopropoxy, tert-butoxy, hydoxymethyl, hydroxyethyl,thiomethyl, thioethyl, aminomethyl, aminoethyl, acetyl, acetoxy,methylamino, ethylamino, dimethylamino, diethylamino,N-methyl-N-ethylamino, acetylamino, N-methylcarbamoyl, N-ethylcarbamoyl,N,N-dimethylcarbamoyl, N,N-diethylcarbamoyl, N-methyl-N-ethylcarbamoyl,methylthio, ethylthio, methylsulfinyl, ethylsulfinyl, mesyl,ethylsulfonyl, methoxycarbonyl, ethoxycarbonyl, isopropoxycarbonyl,tert-butoxycarbonyl, N-methylsulfamoyl, N-ethylsulfamoyl,N,N-dimethylsulfamoyl, N,N-diethylsulfamoyl, N-methyl-N-ethylsulfamoyl,benozyl, benzyl, carbocyclyl, aryl, or heterocyclyl.
 2. The method ofclaim 1, the compound has one of the following formula:

wherein R⁶ is hydrogen, alkyl, or alkanoyl optionally substituted withR¹⁰.
 3. The method of claim 1, the compound has one of the followingformula:

wherein, X is —CH₂—, —C(OH)(OH)—, —C(OH)H, —C(Hal)(Hal)-, —C(Hal)H—,—O—, —S—, —(S═O)—, —SO₂—, —NH—, —(C═O)—, —(C═NH)—, or —(C═S)—; Y is—CH₂—, —C(OH)(OH)—, —C(OH)H, —C(Hal)(Hal)-, —C(Hal)H—, —O—, —S—,—(S═O)—, —SO₂—, —NH—, —(C═O)—, —(C═NH)—, or —(C═S)—; Z is —CH₂—,—C(OH)(OH)—, —C(OH)H, —C(Hal)(Hal)-, —C(Hal)H—, —O—, —S—, —(S═O)—,—SO₂—, —NH—, —(C═O)—, —(C═NH)—, or —(C═S)—; and R¹ is hydrogen, hydroxy,alkyl, alkanoyl, amino, aminoalkyl, carbamoyl, sulfate, sulfonate,aminosulfonyl, phosphate, phosphonate, or heterocyclyl.
 4. The method ofclaim 3, wherein a) X is O, and R¹ is alkanoyl; b) X is —NH—, and R¹ isalkanoyl; c) X is O, and R¹ is aminosulfonyl; d) X is —NH—, and R¹ isaminosulfonyl; e) X is —(C═O)—, R¹ is amino; f) X is O, Y is —(C═O)—, R¹is amino; g) X is O, Y is —(C═O)—, Z is —NH—, and R¹ is sulfonate; andh) X is O, Y is —(C═O)—, Z is —NH—, and R¹ is aminosulfonyl.
 5. Themethod of claim 1, wherein the compound is5-hexyl-4-phenyl-3a-(1-phenylvinyl)-1,2,3,3a,6,6a-hexahydropentalen-1-ylsulfamide or salt thereof.
 6. The method of claim 1, wherein thecompound is5-hexyl-4-phenyl-3a-(1-phenylvinyl)-1,2,3,3a,6,6a-hexahydropentalen-1-yl)acetamideor salt thereof.
 7. The method of claim 1, wherein the cardiovasculardisease is cardiovascular disease is selected from coronary arterydisease (CAD), angina, myocardial infarction, stroke, hypertensive heartdisease, rheumatic heart disease, cardiomyopathy, heart arrhythmia,congenital heart disease, valvular heart disease, carditis, aorticaneurysms, peripheral artery disease, and venous thrombosis.
 8. A methodof treating or preventing diabetes comprising administering to a subjectin need thereof an effective amount of a compound having the followingformula:

prodrugs or salts thereof wherein, R¹ is alkyl, halogen, nitro, cyano,hydroxy, amino, mercapto, formyl, carboxy, carbamoyl, alkoxy,hydroxyalkyl, alkylthio, thioalkyl, alkylamino, aminoalkyl,(alkyl)₂amino, alkanoyl, alkoxycarbonyl, alkylsulfinyl, alkylsulfonyl,arylsulfonyl, carbocyclyl, benozyl, benzyl, aryl, or heterocyclyl,wherein R¹ is optionally substituted with one or more, the same ordifferent, R¹⁰; R² is hydrogen, alkyl, halogen, nitro, cyano, hydroxy,amino, mercapto, formyl, carboxy, carbamoyl, alkoxy, hydroxyalkyl,alkylthio, thioalkyl, alkylamino, aminoalkyl, (alkyl)₂amino, alkanoyl,alkoxycarbonyl, alkylsulfinyl, alkylsulfonyl, arylsulfonyl, carbocyclyl,benozyl, benzyl, aryl, or heterocyclyl, wherein R² is optionallysubstituted with one or more, the same or different, R¹⁰; R³ ishydrogen, alkyl, halogen, nitro, cyano, hydroxy, amino, mercapto,formyl, carboxy, carbamoyl, alkoxy, hydroxyalkyl, alkylthio, thioalkyl,alkylamino, aminoalkyl, (alkyl)₂amino, alkanoyl, alkoxycarbonyl,alkylsulfinyl, alkylsulfonyl, arylsulfonyl, carbocyclyl, benozyl,benzyl, aryl, or heterocyclyl, wherein R³ is optionally substituted withone or more, the same or different, R¹⁰; R⁴ is alkyl, halogen, nitro,cyano, hydroxy, amino, mercapto, formyl, carboxy, carbamoyl, alkoxy,hydroxyalkyl, alkylthio, thioalkyl, alkylamino, aminoalkyl,(alkyl)₂amino, alkanoyl, alkoxycarbonyl, alkylsulfinyl, alkylsulfonyl,arylsulfonyl, carbocyclyl, benozyl, benzyl, aryl, or heterocyclyl,wherein R⁴ is optionally substituted with one or more, the same ordifferent, R¹⁰; R⁵ is alkyl, halogen, nitro, cyano, hydroxy, amino,mercapto, formyl, carboxy, carbamoyl, alkoxy, hydroxyalkyl, alkylthio,thioalkyl, alkylamino, aminoalkyl, (alkyl)₂amino, alkanoyl,alkoxycarbonyl, alkylsulfinyl, alkylsulfonyl, arylsulfonyl, carbocyclyl,benozyl, benzyl, aryl, or heterocyclyl, wherein R⁶ is optionallysubstituted with one or more, the same or different, R¹⁰; R⁶ is lipid oralkyl, wherein R⁶ is optionally terminally substituted with a hydroxy,carboxy, or phosphate, wherein the hydroxy, carboxy, or phosphate areoptionally further substituted with R¹⁰; R⁷ is hydrogen, alkyl, halogen,nitro, cyano, hydroxy, amino, mercapto, formyl, carboxy, carbamoyl,alkoxy, hydroxyalkyl, alkylthio, thioalkyl, alkylamino, aminoalkyl,(alkyl)₂amino, alkanoyl, alkoxycarbonyl, alkylsulfinyl, alkylsulfonyl,arylsulfonyl, carbocyclyl, benozyl, benzyl, aryl, or heterocyclyl,wherein R⁷ is optionally substituted with one or more, the same ordifferent, R¹⁰; or R¹ and R⁷ together are an oxo or oxime, wherein theoxime is optionally substituted with one or more, the same or different,R¹⁰; R¹⁰ is alkyl, halogen, nitro, cyano, hydroxy, amino, mercapto,formyl, carboxy, carbamoyl, alkoxy, hydroxyalkyl, alkylthio, thioalkyl,alkylamino, aminoalkyl, (alkyl)₂amino, alkanoyl, alkoxycarbonyl,alkylsulfinyl, alkylsulfonyl, arylsulfonyl, aminosulfonyl, carbocyclyl,benozyl, benzyl, aryl, or heterocyclyl, wherein R¹⁰ is optionallysubstituted with one or more, the same or different, R¹¹; and R¹¹ ishalogen, nitro, cyano, hydroxy, trifluoromethoxy, trifluoromethyl,amino, formyl, carboxy, carbamoyl, mercapto, sulfamoyl, methyl, ethyl,methoxy, ethoxy, isopropoxy, tert-butoxy, hydoxymethyl, hydroxyethyl,thiomethyl, thioethyl, aminomethyl, aminoethyl, acetyl, acetoxy,methylamino, ethylamino, dimethylamino, diethylamino,N-methyl-N-ethylamino, acetylamino, N-methylcarbamoyl, N-ethylcarbamoyl,N,N-dimethylcarbamoyl, N,N-diethylcarbamoyl, N-methyl-N-ethylcarbamoyl,methylthio, ethylthio, methylsulfinyl, ethylsulfinyl, mesyl,ethylsulfonyl, methoxycarbonyl, ethoxycarbonyl, isopropoxycarbonyl,tert-butoxycarbonyl, N-methylsulfamoyl, N-ethylsulfamoyl,N,N-dimethylsulfamoyl, N,N-diethylsulfamoyl, N-methyl-N-ethylsulfamoyl,benozyl, benzyl, carbocyclyl, aryl, or heterocyclyl.
 9. The method ofclaim 8, the compound has one of the following formula:

wherein R⁶ is hydrogen, alkyl, or alkanoyl optionally substituted withR¹⁰.
 10. The method of claim 8, the compound has one of the followingformula:

wherein, X is —CH₂—, —C(OH)(OH)—, —C(OH)H, —C(Hal)(Hal)-, —C(Hal)H—,—O—, —S—, —(S═O)—, —SO₂—, —NH—, —(C═O)—, —(C═NH)—, or —(C═S)—; Y is—CH₂—, —C(OH)(OH)—, —C(OH)H, —C(Hal)(Hal)-, —C(Hal)H—, —O—, —S—,—(S═O)—, —SO₂—, —NH—, —(C═O)—, —(C═NH)—, or —(C═S)—; Z is —CH₂—,—C(OH)(OH)—, —C(OH)H, —C(Hal)(Hal)-, —C(Hal)H—, —O—, —S—, —(S═O)—,—SO₂—, —NH—, —(C═O)—, —(C═NH)—, or —(C═S)—; and R¹ is hydrogen, hydroxy,alkyl, alkanoyl, amino, aminoalkyl, carbamoyl, sulfate, sulfonate,aminosulfonyl, phosphate, phosphonate, or heterocyclyl.
 11. The methodof claim 10, wherein a) X is O, and R¹ is alkanoyl; b) X is —NH—, and R¹is alkanoyl; c) X is O, and R¹ is aminosulfonyl; d) X is —NH—, and R¹ isaminosulfonyl; e) X is —(C═O)—, R¹ is amino; f) X is O, Y is —(C═O)—, R¹is amino; g) X is O, Y is —(C═O)—, Z is —NH—, and R¹ is sulfonate; andh) X is O, Y is —(C═O)—, Z is —NH—, and R¹ is aminosulfonyl.
 12. Themethod of claim 8, wherein the compound is5-hexyl-4-phenyl-3a-(1-phenylvinyl)-1,2,3,3a,6,6a-hexahydropentalen-1-ylsulfamide or salt thereof.
 13. The method of claim 8, wherein thecompound is5-hexyl-4-phenyl-3a-(1-phenylvinyl)-1,2,3,3a,6,6a-hexahydropentalen-1-yl)acetamideor salt thereof.
 14. The method of claim 8, wherein diabetes is selectedfrom insulin-dependent diabetes mellitus, non insulin-dependent diabetesmellitus, and gestational diabetes.
 15. A method of treating orpreventing cancer comprising administering to a subject in need thereofan effective amount of a compound having the following formula:

prodrugs or salts thereof wherein, R¹ is alkyl, halogen, nitro, cyano,hydroxy, amino, mercapto, formyl, carboxy, carbamoyl, alkoxy,hydroxyalkyl, alkylthio, thioalkyl, alkylamino, aminoalkyl,(alkyl)₂amino, alkanoyl, alkoxycarbonyl, alkylsulfinyl, alkylsulfonyl,arylsulfonyl, carbocyclyl, benozyl, benzyl, aryl, or heterocyclyl,wherein R¹ is optionally substituted with one or more, the same ordifferent, R¹⁰; R² is hydrogen, alkyl, halogen, nitro, cyano, hydroxy,amino, mercapto, formyl, carboxy, carbamoyl, alkoxy, hydroxyalkyl,alkylthio, thioalkyl, alkylamino, aminoalkyl, (alkyl)₂amino, alkanoyl,alkoxycarbonyl, alkylsulfinyl, alkylsulfonyl, arylsulfonyl, carbocyclyl,benozyl, benzyl, aryl, or heterocyclyl, wherein R² is optionallysubstituted with one or more, the same or different, R¹⁰; R³ ishydrogen, alkyl, halogen, nitro, cyano, hydroxy, amino, mercapto,formyl, carboxy, carbamoyl, alkoxy, hydroxyalkyl, alkylthio, thioalkyl,alkylamino, aminoalkyl, (alkyl)₂amino, alkanoyl, alkoxycarbonyl,alkylsulfinyl, alkylsulfonyl, arylsulfonyl, carbocyclyl, benozyl,benzyl, aryl, or heterocyclyl, wherein R³ is optionally substituted withone or more, the same or different, R¹⁰; R⁴ is alkyl, halogen, nitro,cyano, hydroxy, amino, mercapto, formyl, carboxy, carbamoyl, alkoxy,hydroxyalkyl, alkylthio, thioalkyl, alkylamino, aminoalkyl,(alkyl)₂amino, alkanoyl, alkoxycarbonyl, alkylsulfinyl, alkylsulfonyl,arylsulfonyl, carbocyclyl, benozyl, benzyl, aryl, or heterocyclyl,wherein R⁴ is optionally substituted with one or more, the same ordifferent, R¹⁰; R⁵ is alkyl, halogen, nitro, cyano, hydroxy, amino,mercapto, formyl, carboxy, carbamoyl, alkoxy, hydroxyalkyl, alkylthio,thioalkyl, alkylamino, aminoalkyl, (alkyl)₂amino, alkanoyl,alkoxycarbonyl, alkylsulfinyl, alkylsulfonyl, arylsulfonyl, carbocyclyl,benozyl, benzyl, aryl, or heterocyclyl, wherein R⁵ is optionallysubstituted with one or more, the same or different, R¹⁰; R⁶ is lipid oralkyl, wherein R⁶ is optionally terminally substituted with a hydroxy,carboxy, or phosphate, wherein the hydroxy, carboxy, or phosphate areoptionally further substituted with R¹⁰; R⁷ is hydrogen, alkyl, halogen,nitro, cyano, hydroxy, amino, mercapto, formyl, carboxy, carbamoyl,alkoxy, hydroxyalkyl, alkylthio, thioalkyl, alkylamino, aminoalkyl,(alkyl)₂amino, alkanoyl, alkoxycarbonyl, alkylsulfinyl, alkylsulfonyl,arylsulfonyl, carbocyclyl, benozyl, benzyl, aryl, or heterocyclyl,wherein R⁷ is optionally substituted with one or more, the same ordifferent, R¹⁰; or R¹ and R⁷ together are an oxo or oxime, wherein theoxime is optionally substituted with one or more, the same or different,R¹⁰; R¹⁰ is alkyl, halogen, nitro, cyano, hydroxy, amino, mercapto,formyl, carboxy, carbamoyl, alkoxy, hydroxyalkyl, alkylthio, thioalkyl,alkylamino, aminoalkyl, (alkyl)₂amino, alkanoyl, alkoxycarbonyl,alkylsulfinyl, alkylsulfonyl, arylsulfonyl, aminosulfonyl, carbocyclyl,benozyl, benzyl, aryl, or heterocyclyl, wherein R¹⁰ is optionallysubstituted with one or more, the same or different, R¹¹; and R¹¹ ishalogen, nitro, cyano, hydroxy, trifluoromethoxy, trifluoromethyl,amino, formyl, carboxy, carbamoyl, mercapto, sulfamoyl, methyl, ethyl,methoxy, ethoxy, isopropoxy, tert-butoxy, hydoxymethyl, hydroxyethyl,thiomethyl, thioethyl, aminomethyl, aminoethyl, acetyl, acetoxy,methylamino, ethylamino, dimethylamino, diethylamino,N-methyl-N-ethylamino, acetylamino, N-methylcarbamoyl, N-ethylcarbamoyl,N,N-dimethylcarbamoyl, N,N-diethylcarbamoyl, N-methyl-N-ethylcarbamoyl,methylthio, ethylthio, methylsulfinyl, ethylsulfinyl, mesyl,ethylsulfonyl, methoxycarbonyl, ethoxycarbonyl, isopropoxycarbonyl,tert-butoxycarbonyl, N-methylsulfamoyl, N-ethylsulfamoyl,N,N-dimethylsulfamoyl, N,N-diethylsulfamoyl, N-methyl-N-ethylsulfamoyl,benozyl, benzyl, carbocyclyl, aryl, or heterocyclyl.
 16. The method ofclaim 15, the compound has one of the following formula:

wherein R⁶ is hydrogen, alkyl, or alkanoyl optionally substituted withR¹⁰.
 17. The method of claim 15, the compound has one of the followingformula:

wherein, X is —CH₂—, —C(OH)(OH)—, —C(OH)H, —C(Hal)(Hal)-, —C(Hal)H—,—O—, —S—, —(S═O)—, —SO₂—, —NH—, —(C═O)—, —(C═NH)—, or —(C═S)—; Y is—CH₂—, —C(OH)(OH)—, —C(OH)H, —C(Hal)(Hal)-, —C(Hal)H—, —O—, —S—,—(S═O)—, —SO₂—, —NH—, —(C═O)—, —(C═NH)—, or —(C═S)—; Z is —CH₂—,—C(OH)(OH)—, —C(OH)H, —C(Hal)(Hal)-, —C(Hal)H—, —O—, —S—, —(S═O)—,—SO₂—, —NH—, —(C═O)—, —(C═NH)—, or —(C═S)—; and R¹ is hydrogen, hydroxy,alkyl, alkanoyl, amino, aminoalkyl, carbamoyl, sulfate, sulfonate,aminosulfonyl, phosphate, phosphonate, or heterocyclyl.
 18. The methodof claim 17, wherein a) X is O, and R¹ is alkanoyl; b) X is —NH—, and R¹is alkanoyl; c) X is O, and R¹ is aminosulfonyl; d) X is —NH—, and R¹ isaminosulfonyl; e) X is —(C═O)—, R¹ is amino; f) X is O, Y is —(C═O)—, R¹is amino; g) X is O, Y is —(C═O)—, Z is —NH—, and R¹ is sulfonate; andh) X is O, Y is —(C═O)—, Z is —NH—, and R¹ is aminosulfonyl.
 19. Themethod of claim 15, wherein the compound is5-hexyl-4-phenyl-3a-(1-phenylvinyl)-1,2,3,3a,6,6a-hexahydropentalen-1-ylsulfamide or salt thereof.
 20. The method of claim 15, wherein thecompound is5-hexyl-4-phenyl-3a-(1-phenylvinyl)-1,2,3,3a,6,6a-hexahydropentalen-1-yl)acetamideor salt thereof.